U.S. patent application number 10/536606 was filed with the patent office on 2006-08-03 for virulence genes, proteins, and their use.
Invention is credited to Mohammed Benghezal, Pierre Cosson, Christian Van Delden, Thilo Kohler, Anna Marchetti.
Application Number | 20060171959 10/536606 |
Document ID | / |
Family ID | 32593615 |
Filed Date | 2006-08-03 |
United States Patent
Application |
20060171959 |
Kind Code |
A1 |
Cosson; Pierre ; et
al. |
August 3, 2006 |
Virulence genes, proteins, and their use
Abstract
A series of genes from Pseudomonas aeruginosa and Klebsiella are
shown to encode products that are implicated in virulence. The
identification of theses genes therefore allows attenuated
microorganisms to be produced. Furthermore, the genes or their
encoded products can be used to identify antimicrobial drugs,
diagnostic methods for the identification of a pathogen-associated
disease, and in the manufacture of vaccines.
Inventors: |
Cosson; Pierre; (Carouge,
CH) ; Kohler; Thilo; (Carouge, CH) ;
Benghezal; Mohammed; (Grand-Saconnex, CH) ;
Marchetti; Anna; (Geneva, CH) ; Delden; Christian
Van; (Colony, CH) |
Correspondence
Address: |
MINTZ, LEVIN, COHN, FERRIS, GLOVSKY;AND POPEO, P.C.
ONE FINANCIAL CENTER
BOSTON
MA
02111
US
|
Family ID: |
32593615 |
Appl. No.: |
10/536606 |
Filed: |
December 19, 2003 |
PCT Filed: |
December 19, 2003 |
PCT NO: |
PCT/CH03/00836 |
371 Date: |
December 15, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
10324967 |
Dec 20, 2002 |
6974680 |
|
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10536606 |
Dec 15, 2005 |
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Current U.S.
Class: |
424/200.1 ;
424/260.1; 435/252.34 |
Current CPC
Class: |
A61K 2039/522 20130101;
C07K 14/26 20130101; C07K 14/21 20130101 |
Class at
Publication: |
424/200.1 ;
424/260.1; 435/252.34 |
International
Class: |
A61K 39/02 20060101
A61K039/02; A61K 39/108 20060101 A61K039/108; C12N 1/21 20060101
C12N001/21 |
Claims
1. An attenuated bacterial mutant derived from a pathogenic
bacterial strain, wherein said attenuated mutant has: (i) a
mutation of a gene selected from the group consisting of VIR1,
VIR2, VIR3, VIR4, VIR5, VIR6, VIR7, VIR8, VIR9, VIR10, VIR11,
VIR12, VIR13, VIR14. VIR15, VIR16, VIR17, VIR18, VIR19, VIR20,
VIR21, VIR22, VIR23, VIR24, VIR25, VIR26, VIR27, VIR28, VIR29,
VIR30, VIR31, VIR32, VIR33, VIR34, VIR35, VIR36, VIR37, VIR38,
VIR39, VIR40, VIR41, VIR42, VIR43, VIR44, VIR45, and VIR46; and
(ii) reduced inhibition of Dictyostelium amoeba growth when
compared to the growth observed in the presence of an isogenic
bacterial strain.
2. An attenuated bacterial mutant of claim 1, wherein said mutation
is insertional inactivation or a gene deletion.
3. An attenuated bacterial mutant of claim 1, wherein said mutant
is a gram-negative bacteria.
4. An attenuated bacterial mutant of claim 3, wherein said
attenuated gram-negative bacterial mutant is a Pseudomonas
species.
5. An attenuated bacterial mutant of claim 4, wherein said
Pseudomonas species is Pseudomonas aeruginosa.
6. An attenuated Pseudomonas mutant of claim 5, wherein said
attenuated Pseudomonas mutant is selected from the group consisting
of: MUT1; MUT2; MUT3; MUT4; MUT5; MUT6; MUT7; MUT8; MUT9; MUT10;
MUT11; MUT12; MUT13; MUT14; MUT15; MUT16; MUT17; MUT18; and
MUT19.
7. An attenuated bacterial mutant of claim 3, wherein said
gram-negative bacterial mutant is a Klebsiella species.
8. An attenuated bacterial mutant of claim 7, wherein said
Klebsiella species is Klebsiella pneumoniae.
9. An attenuated Klebsiella mutant of claim 8, wherein said
attenuated Klebsiella mutant is selected from the group consisting
of: MUT20; MUT21; MUT22; MUT23; MUT24; MUT25; MUT26; MUT27; MUT28;
MUT29; MUT30; MUT31; MUT32; MUT33; MUT34; MUT35; MUT36; MUT37;
MUT38; MUT39; MUT40; MUT41; MUT42; MUT43; MUT44; MUT45; and
MUT46.
10. A method for identifying an antimicrobial drug, said method
comprising: (a) contacting a candidate composition with at least
one polypeptide encoded by a gene selected from the group
consisting of VIR1, VIR2, VIR3, VIR4, VIR5, VIR6, VIR7, VIR8, VIR9,
VIR10, VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17, VIR18,
VIR19, VIR20, VIR21, VIR22, VIR23, VIR24, VIR25, VIR26, VIR27,
VIR28, VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35, VIR36,
VIR37, VIR38, VIR39, VIR40, VIR41, VIR42, VIR43, VIR44, VIR45 and
VIR46; and (b) comparing the biological activity of said
polypeptide in the presence and absence of said candidate
composition, wherein alteration of the biological activity of said
polypeptide indicates that said candidate composition is an
antimicrobial drug.
11. A method of claim 10, wherein said candidate composition
contains at least two molecules.
12. A method of claim 10, wherein said candidate composition
contains at least one-molecule less than about 500 Daltons.
13. A method of claim 10, wherein said candidate composition
contains at least one molecule greater than about 500 Daltons.
14. A method of claim 10, wherein said candidate composition
contains at least one molecule selected from a group consisting of
a polypeptide, polysaccharide, lipid, nucleic acid, or combination
thereof.
15. A composition of claim 14, wherein said polypeptide is an
immunoglobulin.
16. A method for identifying an antimicrobial drug, said method
comprising: (a) contacting at a candidate composition with at least
one polynucleotide encoded by a gene selected from the group
consisting of VIR1, VIR2, VIR3, VIR4, VIR5, VIR6, VIR7, VIR8, VIR9,
VIR10, VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17, VIR18,
VIR19, VIR20, VIR21, VIR22, VIR23, VIR24, VIR25, VIR26, VIR27,
VIR28, VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35, VIR36,
VIR37, VIR38, VIR39, VIR40, VIR41, VIR42, VIR43, VIR44, VIR45, and
VIR46; and (b) comparing the expression of said polynucleotide in
the presence and absence of said candidate composition, wherein
alteration of the expression of said nucleotide indicates that said
candidate composition is an antimicrobial drug.
17. A method of claim 16, wherein said candidate composition
contains at least two molecules.
18. A method of claim 16, wherein said candidate composition
contains at least one molecule less than about 500 Daltons.
19. A method of claim 16, wherein said candidate composition
contains at least one molecule greater than about 500 Daltons.
20. A method of claim 16, wherein said candidate composition
contains at least one molecule selected from a group consisting of
a polypeptide, polysaccharide, lipid, nucleic acid, or combination
thereof.
21. A composition of claim 20, wherein said nucleic acid is a
ribonucleic acid.
22. A nucleic acid of claim 21, wherein said nucleic acid is a
small interfering ribonucleic acid.
23. A method for determining the degree of virulence of a pathogen
in a subject, said method comprising: (a) measuring the level of
expression of at least one polypeptide encoded by a gene selected
from the group consisting of VIR1, VIR2, VIR3, VIR4, VIR5, VIR6,
VIR7, VIR8, VIR9, VIR10, VIR11, VIR12, VIR13, VIR14, VIR15, VIR16,
VIR17, VIR18, VIR19, VIR20, VIR21, VIR22, VIR23, VIR24, VIR25,
VIR26, VIR27, VIR28, VIR29, VIR30, VIR31, VIR32, VIR33, VIR34,
VIR35, VIR36, VIR37, VIR38, VIR39, VIR40, VIR41, VIR42, VIR43,
VIR44, VIR45, and VIR46, in a sample from the first subject; and
(b) comparing the amount of said polypeptide in said sample of step
(a) to the amount of said polypeptide present in a control sample
from a second subject known not to have the presence of said
pathogen, wherein an alteration in the expression level of said
polypeptide in said first subject as compared to said control
sample indicates the degree of virulence of said pathogen.
24. A method of claim 23, wherein said subject is a mammal.
25. A mammalian subject of claim 24, wherein said mammalian subject
is a human.
26. A method for determining the degree of virulence of a pathogen
in a subject, said method comprising: (a) measuring the level of
expression of at least one polynucleotide encoded by a gene
selected from the group consisting of VIR1, VIR2, VIR3, VIR4, VIR5,
VIR6, VIR7, VIR8, VIR9, VIR10, VIR11, VIR12, VIR13, VIR14, VIR15,
VIR16, VIR17, VIR18, VIR19, VIR20, VIR21, VIR22, VIR23, VIR24,
VIR25, VIR26, VIR27, VIR28, VIR29, VIR30, VIR31, VIR32, VIR33,
VIR34, VIR35, VIR36, VIR37, VIR38, VIR39, VIR40, VIR41, VIR42,
VIR44, VIR45, and VIR46, in a sample from the first subject; and
(b) comparing the amount of said polynucleotide in said sample of
step (a) to the amount of said polynucleotide present in a control
sample from a second subject known not to have the presence of said
pathogen, wherein an alteration in the expression level of said
polynucleotide in said first subject as compared to said control
sample indicates the degree of virulence of said pathogen.
27. A method of claim 26, wherein said subject is a mammal.
28. A mammalian subject of claim 27, wherein said mammalian subject
is a human.
29. An attenuated bacterial mutant of claim 1, wherein said mutant
encodes and expresses a foreign antigen.
30. An attenuated bacterial mutant of claim 1, wherein said mutant
contains a plasmid which encodes and expresses, in a eukaryotic
cell, a foreign antigen.
31. A vaccine against a disease caused by a pathogenic
microorganism comprising: (a) a pharmaceutically effective dosage
of one or more of the attenuated bacterial mutants of claim 1 and;
(b) a pharmaceutically acceptable diluent or carrier.
32. An attenuated bacterial mutant derived from a pathogenic
bacterial strain, wherein said attenuated mutant has: (i) a
mutation of a gene selected from the group consisting of pchE,
pchF, pchG, pchH, and pchI; and (ii) reduced inhibition of
Dictyostelium amoeba growth when compared to the growth observed in
the presence of an isogenic bacterial strain.
33. A bacterial strain comprising an operon encoding a gene
selected from the group consisting of VIR1, VIR2, VIR3, VIR4, VIR5,
VIR6, VIR7, VIR8, VIR9, VIR10, VIR11, VIR12, VIR13, VIR14, VIR15,
VIR16, VIR17, VIR18, VIR19, VIR20, VIR21, VIR22, VIR23, VIR24,
VIR25, VIR26, VIR27, VIR28, VIR29, VIR30, VIR31, VIR32, VIR33,
VIR34, VIR35, VIR36, VIR37, VIR38, VIR39, VIR40, VIR41, VIR42,
VIR44, VIR45, and VIR46, wherein said bacterial strain includes a
mutation that reduces expression of said gene relative to an
isogenic bacterial strain lacking said mutation.
34. A bacterial strain of claim 33, wherein said mutation reduces
inhibition of Dictyostelium amoeba growth when compared to the
growth of Dictyostelium amoeba in the presence of an isogenic
bacterial strain lacking said mutation.
Description
FIELD OF THE INVENTION
[0001] This invention relates to virulence genes and proteins, and
their use. More particularly, it relates to genes and
proteins/peptides obtained from gram-negative bacteria, and their
use in therapy and in screening for drugs.
BACKGROUND OF THE INVENTION
[0002] According to health care experts, infectious diseases caused
by microbes are responsible for more deaths worldwide than any
other single cause. The current estimate of the annual cost of
medical care for treating infectious diseases in the United States
alone is about $120 billion. While antibiotic treatment is
effective for many microbial infections, antibiotic resistance
among pathogenic bacteria is a growing health concern. Indeed, the
American Medical Association has concluded that, "the global
increase in resistance to antimicrobial drugs, including the
emergence of bacterial strains that are resistant to all available
antibacterial agents, has created a public health problem of
potentially crisis proportions."
[0003] Pseudomonas and Klebsiella are two genuses of gram-negative
bacteria that pose a significant health risk to infected host
organisms, in part, due to their resistance to many antibiotics.
These bacteria are noted for causing life-threatening infections,
particularly in the lung. Cancer and burn patients also commonly
suffer serious Pseudomonas infections, as do certain other
individuals with immune system deficiencies. While Klebsiella sp.
is responsible for many types of infections, outside of a medical
setting, the most common infection caused by Klebsiella bacteria is
pneumonia.
[0004] There is a need in the art for new antimicrobial therapeutic
strategies.
SUMMARY OF THE INVENTION
[0005] The present invention is based, in part, on the discovery of
46 genes, when mutated lower the virulence of a gram-negative
bacterium, and can be used in new antimicrobial therapeutic
strategies. The invention provides attenuated bacterial mutants
that are derived from pathogenic strains. These attenuated
bacterial stains have a mutation in a VIRX gene identified herein
as VIR1, VIR2, VIR3, VIR4, VIR5, VIR6, VIR7, VIR8, VIR9, VIR10,
VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17, VIR18, VIR19,
VIR20, VIR21, VIR22, VIR23, VIR24, VIR25, VIR26, VIR27, VIR28,
VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35, VIR36, VIR37,
VIR38, VIR39, VIR40, VIR41, VIR42, VIR43, VIR44, VIR45, and VIR46;
and show reduced inhibition of Dictyostelium amoeba growth when
compared to the growth observed in the presence of an isogenic
bacterial strain. The term, "pathogenic," as used herein, is
defined as an agent's ability to cause disease, damage or harm to a
host organism. The term, "attenuated," as used herein, means an
organism made less virulent relative to an isogenic pathogenic
organism. The term, "mutant," as used herein, an organism carrying
a specific mutation of a gene that is expressed in the organism's
phenotype. A mutation may be insertional inactivation or deletion
of a gene. It is preferred that the mutation be an insertional
inactivation of a gene.
[0006] The invention also provides attenuated bacterial mutants
that are derived from pathogenic gram-negative bacterial strains.
These attenuated gram-negative bacterial strains have a mutation in
a VIRX gene identified herein as VIR1, VIR2, VIR3, VIR4, VIR5,
VIR6, VIR7, VIR8, VIR9, VIR10, VIR11, VIR12, VIR13, VIR14, VIR15,
VIR16, VIR17, VIR18, VIR19, VIR20, VIR21, VIR22, VIR23, VIR24,
VIR25, VIR26, VIR27, VIR28, VIR29, VIR30, VIR31, VIR32, VIR33,
VIR34, VIR35, VIR36, VIR37, VIR38, VIR39, VIR40, VIR41, VIR42,
VIR43, VIR44, VIR45, and VIR46; and show reduced inhibition of
Dictyostelium amoeba growth when compared to the growth observed in
the presence of an isogenic bacterial strain. A mutation may be
insertional inactivation or deletion of a gene. It is preferred
that the mutation be an insertional inactivation of a gene. It is
also preferred that the attenuated gram-negative bacterial mutant
be derived from a Pseudomonas or Klebiella spp. It is more
preferred that the attenuated gram-negative bacterial mutant is a
strain of P. aeruginosa or K. pneumoniae.
[0007] The invention additionally provides for a VIRX gene that may
be part of an operon. The term, "operon," as used herein, is a unit
of bacterial gene expression and regulation comprising several
genes, usually with complementary functions. Insertion in a gene in
an operon typically interferes with the function of this gene and
of other genes located downstream or upstream in the operon. The
function attributed to a gene refers to its function and/or that of
any gene located downstream or upstream in the same operon.
Accordingly, the invention also provides for a bacterial strain
comprising an operon encoding a gene selected from the group
consisting of VIR1, VIR2, VIR3, VIR4, VIR5, VIR6, VIR7, VIR8, VIR9,
VIR10, VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17, VIR18,
VIR19, VIR20, VIR21, VIR22, VIR23, VIR24, VIR25, VIR26, VIR27,
VIR28, VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35, VIR36,
VIR37, VIR38, VIR39, VIR40, VIR41, VIR42, VIR44, VIR45, and VIR46,
wherein the bacterial strain includes a mutation that reduces
expression of the VIRX gene relative to an isogenic bacterial
strain lacking the mutation. In one embodiment, the the mutation
reduces inhibition of Dictyostelium amoeba growth when compared to
the growth of Dictyostelium amoeba in the presence of an isogenic
bacterial strain lacking the mutation.
[0008] The invention provides for one or more of the following
attenuated Pseudomonas mutant strains: MUT1; MUT2; MUT3; MUT4;
MUT5; MUT6; MUT7; MUT8; MUT9; MUT10; MUT11; MUT12; MUT13; MUT14;
MUT15: MUT16; MUT17; MUT18; and MUT19. The invention also provides
for one or more of the following attenuated Klebsiella mutant
strains: MUT20; MUT21; MUT22; MUT23; MUT24; MUT25; MUT26; MUT27;
MUT28; MUT29; MUT30; MUT31; MUT32; MUT33; MUT34; MUT35; MUT36;
MUT37; MUT38; MUT39; MUT40; MUT41; MUT42; MUT43; MUT44; MUT45; and
MUT46.
[0009] The invention additionally provides a method for identifying
an antimicrobial drug, wherein a candidate composition is contacted
with at least one polypeptide encoded by a gene selected from the
group consisting of VIR1, VIR2, VIR3, VIR4, VIR5, VIR6, VIR7, VIR8,
VIR9, VIR10, VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17,
VIR18, VIR19, VIR20, VIR21, VIR22, VIR23, VIR24, VIR25, VIR26,
VIR27, VIR28, VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35,
VIR36, VIR37, VIR38, VIR39, VIR40, VIR41, VIR42, VIR43, VIR44,
VIR45 and VIR46. The biological activity of polypeptide in the
presence of the candidate composition is compared with the
biological activity of the polypeptide in the absence of the
candidate composition. Alteration of the biological activity of the
polypeptide indicates that the candidate composition is an
antimicrobial drug. In some embodiments, the candidate composition
contains at least two molecules. The candidate composition can
contain at least one molecule less than about 500 Daltons or at
least one molecule greater than about 500 Daltons. The candidate
composition can be, e.g., an immunoglobulin, polysaccharide, lipid,
nucleic acid, or combination thereof.
[0010] The invention additionally provides a method for identifying
an antimicrobial drug, wherein a candidate composition is contacted
with at least one polynucleotide encoded by a gene selected from
the group consisting of VIR1, VIR2, VIR3, VIR4, VIR5, VIR6, VIR7,
VIR8, VIR9, VIR10, VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17,
VIR18, VIR19, VIR20, VIR21, VIR22, VIR23, VIR24, VIR25, VIR26,
VIR27, VIR28, VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35,
VIR36, VIR37, VIR38, VIR39, VIR40, VIR41, VIR42, VIR43, VIR44,
VIR45, and VIR46. The expression of the polynucleotide in the
presence of the candidate composition is compared with the
expression of the polynucleotide in the absence of the candidate
composition. Alteration of the expression of the polynucleotide
indicates that the candidate composition is an antimicrobial drug.
In some embodiments, the candidate composition contains at least
two molecules. The candidate composition can contain at least one
molecule less than about 500 Daltons or at least one molecule
greater than about 500 Daltons. The candidate composition can be a
polypeptide, polysaccharide, lipid, nucleic acid, e.g., ribonucleic
acid, or combination thereof. In a preferred embodiment, the
ribonucleic acid of the candidate composition is a small
interfering ribonucleic acid.
[0011] The invention additionally provides a method for determining
the degree of virulence of a pathogen present in a subject,
comprising: [0012] (a) measuring the level of expression of at
least one polypeptide encoded by a gene selected from the group
consisting of VIR1, VIR2, VIR3, VIR4, VIR5, VIR6, VIR7, VIR8, VIR9,
VIR10, VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17, VIR18,
VIR19, VIR20, VIR21, VIR22, VIR23, VIR24, VIR25, VIR26, VIR27,
VIR28, VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35, VIR36,
VIR37, VIR38, VIR39, VIR40, VIR41, VIR42, VIR43, VIR44, VIR45, and
VIR46, in a sample from the first subject; and [0013] (b) comparing
the amount of the polypeptide in the sample of step (a) to the
amount of the polypeptide present in a control sample from a second
subject known not to have the presence of the pathogen, wherein an
alteration in the expression level of the polypeptide in the first
subject as compared to the control sample indicates the degree of
virulence of the pathogen.
[0014] In a preferred embodiment, the subject is a mammal. It is
more preferred that the subject is a human.
[0015] The invention also provides a method for determining the
degree of virulence of a pathogen present in a subject,
comprising:
[0016] (a) measuring the level of expression of at least one
polynucleotide encoded by a gene selected from the group consisting
of VIR1, VIR2, VIR3, VIR4, VIR5, VIR6, VIR7, VIR8, VIR9, VIR10,
VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17, VIR18, VIR19,
VIR20, VIR21, VIR22, VIR23, VIR24, VIR5, VIR26, VIR27, VIR28,
VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35, VIR36, VIR37,
VIR38, VIR39, VIR40, VIR41, VIR42, VIR44, VIR45, and VIR46, in a
sample from the first subject; and
[0017] (b) comparing the amount of the polynucleotide in the sample
of step (a) to the amount of the polynucleotide present in a
control sample from a second subject known not to have the presence
of the pathogen, wherein an alteration in the expression level of
the polypeptide in the first subject as compared to the control
sample indicates the degree of virulence of the pathogen.
[0018] In a preferred embodiment, the subject is a mammal. It is
more preferred that the subject is a human.
[0019] The invention additionally provides attenuated bacterial
strains that can be used as vaccines and as vectors for foreign
antigens and for foreign DNA. These attenuated bacterial strains
are useful for the preparation of vaccines effective against
diseases associated with the corresponding bacterial strains. In a
preferred embodiment, the attenuated bacterial strains are derived
from Pseudomonas or Klebsiella spp.
[0020] The invention additionally provides attenuated bacterial
strains that can be used as vectors for foreign genes cloned from
other pathogens that will be expressed into proteins, and will
raise protective immune responses against the pathogens from which
they are derived. In a preferred embodiment, the attenuated
bacterial strains used as the vectors are derived from Pseudomonas
or Klebsiella spp.
[0021] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. Although
methods and materials similar or equivalent to those described
herein can be used in the practice or testing of the invention,
suitable methods and materials are described below. All
publications, patent applications, patents, and other references
mentioned herein are incorporated by reference in their entirety.
In the case of conflict, the present specification, including
definitions, will control. In addition, the materials, methods, and
examples are illustrative only and not intended to be limiting.
[0022] Other features and advantages of the invention will be
apparent from the following detailed description and claims.
DETAILED DESCRIPTION OF THE INVENTION
[0023] The present invention is based, in part, on the discovery of
46 genes when mutated lower the virulence of a gram-negative
bacterium. Nineteen of these virulence genes were identified in P.
aeruginosa PT894, while the remaining 27 genes were derived from
mutagenesis of Klebsiella. These bacterial mutants have attenuated
virulence relative to isogenic bacterial strains and are designated
"MUX." Provided herein are virulence genes affected in each novel,
attenuated MUTX strain, as well as the nucleotides and polypeptides
encoded thereby. The sequences encoded by the affected genes are
collectively referred to as "VIRX nucleic acids" or "VIRX
polynucleotides" and the corresponding encoded polypeptides are
referred to as "VIRX polypeptides" or "VIRX proteins." Unless
indicated otherwise, "VIRX" is meant to refer to any of the novel
sequences disclosed herein.
[0024] The peptides and genes of the invention are useful for the
preparation of therapeutic agents to treat infection because they
attenuate the virulence of the wild-type pathogen. Therapy can be
preventative or therapeutic. A subject receiving therapy can be,
e.g., a human, a non-human primate (such as an ape, gorilla, or
chimpanzee), cow, horse, pig, sheep, dog, cat, or rodent (including
mouse or rat).
I. Identification of Pseudomonas and Klebsiella Genes Encoding
Virulence Factors
[0025] Genes encoding virulence factors (e.g., pathogens or toxins)
to a host organism were identified by comparing the growth of
Dictyostelium discoideum, in the presence and absence of test
mutants of Pseudomonas and Klebsiella with an identifiable genetic
alteration as detailed in International Application PCT/IB02/03277,
filed Jun. 7, 2002. Dictyostelium amoebae feed phagocytically upon
bacteria such as K. pneumoniae. When Dictyostelium cells are plated
with K. pneumoniae bacteria, each amoeba creates a plaque in the
bacterial lawn in the region where bacteria have been phagocytosed.
Addition of pathogenic bacteria, e.g., P. aeruginosa strain PT894
to the lawn of K. pneumoniae bacteria, inhibits the growth of the
amoebae.
[0026] Pseudomonas test mutants were made by transposon insertion
according to known methods in the art and tested for virulence in a
Dictyostelium growth assay (see, PCT/IB02/03277, filed Jun. 7,
2002). Klebsiella mutants were also made by transposon insertion
according to known methods in the art and tested for virulence in a
Dictyostelium growth assay (see, PCT/IB02/03277, filed Jun. 7,
2002) using the PHG1a mutant Dictyostelium strain (Cornillon et
al., J. Biol. Chem., 275(44): 34287-92, 2000), a strain which was
found to be particularly sensitive to virulent bacteria.
Specifically, the Klebsiella mutants were obtained by standard
bacteria electroporation technique using the plasposon pNKBOR
(Genbank accession number: AF310136) and selected on solid LB
medium containing 50 .mu.g/ml kanamycin (Rossignol et al., Res.
Microbiol., 152(5): 481-5, 2001). Other mutagenesis methods known
in the art, e.g., ultraviolet radiation exposure, treatment with
intercalating agent or transducing phage, may also be used to
generate mutants. Mutations yielding reduced virulence were
identified where the growth of the Dictyostelium test host organism
exposed to the mutant pathogen was greater than the Dictyostelium
test host organism exposed to wild-type pathogen. Specific genetic
mutations in pathogens displaying reduced virulence were
subsequently identified and characterized by techniques well known
in the art. Identification of specific gene mutations in Klebsiella
mutants was performed by plasmid rescue and cloning of the genomic
DNA at the insertion site mutant using the BglII or ApaI
restriction enzyme according to (Rossignol et al., Res. Microbiol.,
152(5): 481-5, 2001). Identification of specific gene mutations in
Pseudomonas mutants was performed by subcloning the transposon and
surrounding bacteria genomic DNA into an acceptor plamid. DNA
sequencing was performed on amplified rescued plasmids, in order to
identify the insertion site of the transposon. Rat mortality assays
such as that described by Join-Lambert et al., Antimicrob. Agents
Chemother., 45(2): 571-6, 2001, can be used to corroborate
attenuated virulence activity in a mammalian host.
[0027] The 19 Pseudomonas attenuated MUTX organisms harboring the
VIRX genes are summarized below in Table 1. TABLE-US-00001 TABLE 1
STRAIN AFFECTED VIRULENCE GENE(S) REFERENCE MUT1 anthranilate
phosphoribosyltransferase Essar et al, J. Bacteriol., 172: 853-66,
(trpD; PA0650) 1990; Essar et al., J. Bacteriol., 172: 867- 83,
1990. MUT2 ATP sulfurylase small subunit Leyh et al., J. Biol.
Chem., 263: 2409- (CysD; PA4443) 16, 1988; Hummerjohann et al.,
Microbiology, 144 (Pt 5): 1375-86, 1998 MUT3 CysQ (PA5175) Peng and
Verma, J. Biol. Chem., 270: 29105-10, 1995; Neuwald et al., J.
Bacteriol., 174: 415-25, 1992. MUT4 D-amino acid dehydrogenase,
small subunit Lobacka et al., J. Bacteriol., 176: 1500-10, (dadA;
PA5304) 1994. MUT5 imidazoleglycerol-phosphate synthase, cyclase
Fani et al., Mol. Gen. Genet., 216: 224-9, subunit (hisF1; PA5140)
1989; Fani et al., Mol. Gen. Genet., 216: 224-9, 1989. MUT6
N-acetyl-.gamma.-glutamyl-phosphate reductase Smith et al., Gene,
49: 53-60. 1986. (ArgC; PAO 0662) MUT7 Dihydrolipoamide
acetyltransferase (AceF; Rae et al., J. Bacteriol., 179: 3561-71,
pyruvate dehydrogenase complex component 1997. E2; PA5016) MUT8
NADH dehydrogenase I chain H Weidner et al., J. Mol. Biol., 5: 233:
109- (nuoH; PA2643) 22, 1993; Weidner et al., J. Mol. Biol., 233:
109-22, 1993. MUT9 pyoverdine synthetase D Rombel et al., Mol. Gen.
Genet., 246: 519- (PvdD; PA2399) 28, 1995; Merriman et al., J.
Bacteriol., 177: 252-8, 1995. MUT10 RND multidrug efflux
transporter MexD Poole et al., Mol. Microbiol., 21:713-24, (mexD;
PA4598) 1996; Poole et al., Mol. Microbiol., 21: 713-24, 1996.
MUT11 PA3721 Stover et al., Nature, 406: 959-964, 2000. MUT12
PA0596 Tan et al., Proc. Natl. Acad. Sci. USA, 96: 2408-13, 1999.
MUT13 PA5265 Stover et al., Nature, 406: 959-964, 2000. MUT14
pyochelin biosynthetic protein pchC Serino et al., Mol. Gen.
Genet., 249: (PA4229) 217-28, 1995; Serino et al., J. Bactiol.,
179: 248-57, 1997 MUT15 dihydroaeruginoic acid synthetase Reimmann
et al., Microbiology, 144: (pchE; PA4226) 3135-48, 1998. MUT16
Pyochelin synthetase Reimmann et al., Microbiology, 144: (pchF;
PA4225) 3135-48, 1998. MUT17 ATP-binding component of the ABC
Featherston et al., Mol. Microbiol., transporter 32(2): 289-99,
1999; Reimmann et al., J. (pchH; PA4223) Bacteriol., 183: 813-20,
2001. MUT18 ATP-binding component of the ABC Reimmann et al., J.
Bacteriol., 183: 813-20, transporter (pchI; PA4222) 2001. MUT19
putative O-antigen biosynthesis gene cluster Rocchetta et al.,
Microbiol. Mol. Biol. Rev. 63: 523-53, 1999.
[0028] The 27 Klebsiella attenuated MUTX organisms harboring the
VIRX genes disclosed in the present invention and assigned a new
role in virulence are summarized below in Table 2. TABLE-US-00002
TABLE 2 STRAIN AFFECTED VIRULENCE GENE(S) MUT20 hypothetical
transcriptional regulator in met G-dld intergenic region MUT21
.beta.-cystathionase MUT22 ribosome binding factor A MUT23
aspartokinase/homoserine dehydrogenase MUT24 cystathionine
.gamma.-synthase MUT25 Phophoribosylformylglycinamidine synthase
MUT26 homoserine transsuccinylase MUT27 3'-phosphoadenosine
5'-phosphosulfate reductase MUT28 Sfi protein MUT29 transcriptional
activator protein LysR MUT30 TrpD MUT31
N-acetylglucosamine-6-phosphate deacetylase MUT32 WaaQ MUT33
2-Isopropylmalate synthase MUT34 histidinol dehydrogenase MUT35
UDP-galactopyranose mutase MUT36 O-antigen export system permease
protein rfba MUT37 uridyltransferase MUT38 pyridoxine phosphate
biosynthetic protein PdxJ-PdxA MUT39 triose phosphate isomerase
MUT40 aldehyde dehydrogenase MUT41 galactosyl transferase MUT42
siroheme synthetase MUT43
7,8-dihydro-6-hydroxymethylpterin-pyrophosphokinase MUT44
glucose-6-phosphate isomerase MUT45 DNA methylase MUT46 putative
inner membrane protein
II. Attenuated Bacterial Mutants
[0029] A. Attenuated Pseudomonas aeruginosa Mutants
MUT1
[0030] A Pseudomonas bacterial mutant (MUT1) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding anthranilate
phosphoribosyltransferase (PA0650). This gene encodes the VIR1
nucleic acid (SEQ ID NO:1) shown in Table 3A. TABLE-US-00003 TABLE
3A VIR1 Nucleotide Sequence (SEQ ID NO:1)
ATGGATATCAAGGGAGCCCTCAATCGCATCGTCAACCAGCTCGACCTGACCACCGAGGAAATGCAGG
CGGTCATGCGCCAGATCATGACCGGGCAGTGCACCGACGCGCAGATCGGCGCCTTCCTGATGGGCAT
GCGGATGAAGAGCGAAACCATCGACGAGATCGTCGGCGCGGTGGCGGTGATGCGCGAACTGGCCGAC
GGCGTGCAGTTGCCTACGCTGAAGCATGTGGTCGACGTGGTCGGCACCGGCGGCGATGGCGCGAACA
TCTTCAACGTGTCCTCGGCGGCGTCCTTCGTGGTCGCCGCCGCTGGCGGCAAGGTCGCCAAACACGG
TAACCGCGCGGTCTCCGGCAAGAGCGGCAGCGCCGACTTGCTGGAAGCCGCCGGCATCTACCTGGAG
CTGACCTCCGAACAGGTGGCGCGTTGCATCGACACCGTCGGCGTCGGGTTCATGTTCGCCCAGGTCC
ACCACAAGGCGATGAACTACGCCGCCGGTCCGCGCCGCGAGCTGGGCTTGCGGACTCTGTTCAACAT
GCTTGGCCCACTGACCAACCCGGCGGGACTGAGGCACCAGGTGGTCGGGGTGTTCACCCAGGAACTG
TGCAAGCCGCTGGCTGAAGTGCTCAAGCGTCTCGGCAGCGAGCATGTGCTGGTGGTGCATTCGCGCG
ACGGGCTGGACGAGTTCAGTCTGGCCGCGGCGACCCACATTGCCGAGTTGAAGGACGGCGAGGTACG
CGAGTACGAAGTGCGTCCCGAGGACTTCGGGATCAAGAGCCAGACCCTGATGGGGCTGGAGGTCGAC
AGTCCGCAGGCCTCGCTGGAACTGATCCGCGACGCTTTGGGGCGGCGCAAGACCGAGGCTGGGCAGA
AGGCCGCCGAGCTGATCGTGATGAATGCCGGCCCGGCACTGTACGCTGCCGATCTGGCGACCAGCCT
GCACGAGGGCATTCAACTGGCCCACGATGCCCTGCACACCGGGCTGGCACGGGAGAAGATGGACGAA
CTGGTGGCCTTCACCGCCGTTTACAGAGAGGAGAACGCACAGTGA
[0031] The VIR1 protein (SEQ ID NO:2) encoded by SEQ ID NO:1 is
presented using the one-letter amino acid code in Table 3B.
TABLE-US-00004 TABLE 3B Encoded VIR1 protein sequence (SEQ ID NO:2)
MDIKGALNRIVNQLDLTTEEMQAVMRQIMTGQCTDAQIGAFLMGMRMKSETIDEIVGAVAVMREL
ADGVQLPTLKHVVDVVGTGGDGANIFNVSSAASFVVAAAGGKVAKHGNRAVSGKSGSADLLEAAG
IYLELTSEQVARCIDTVGVGFMFAQVHHKAMKYAAGPRRELGLRTLFNMLGPLTNPAGVRHQVVG
VFTQELCKPLAEVLKRLGSEHVLVVHSRDGLDEFSLAAATHIAELKDGEVREYEVRPEDFGIKSQ
TLMGLEVDSPQASLELIRDALGRRKTEAGQKAAELIVMNAGPALYAADLATSLHEGIQLAHDALH
TGLAREKMDELVAFTAVYREENAQ
[0032] The role of VIR1 in virulence was confirmed using phage to
retransduce this mutation into the wild-type PT894 strain where
attenuated virulence was again observed in the Dictyostelium growth
assay compared to an isogenic bacterial strain.
MUT2
[0033] A Pseudomonas bacterial mutant (MUT2) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding the ATP sulfurylase small
subunit (CysD; PA4443). This gene encodes the VIR2 nucleic acid
(SEQ ID NO:3) shown in Table 4A. TABLE-US-00005 TABLE 4A VIR2
Nucleotide Sequence (SEQ ID NO:3)
ATGGTCGACAAACTGACGCACCTGAAACAGCTGGAGGCGGAAAGCATCCA
CATCATCCGCGAGGTGGCCGCCGAGTTCGATAACCCGGTGATGCTGTACT
CGATCGGCAAGGATTCCGCGGTCATGCTGCACCTGGCCCGCAAGGCCTTC
TTCCCCGGCAAGCTGCCCTTCCCGGTGATGCACGTGGACACCCGCTGGAA
ATTCCAGGAGATGTACAGGTTCCGTGATCGGATGGTCGAGGAAATGGGCC
TGGATCTGATCACCCACGTCAACCCGGACGGCGTCGCCCAGGGCATCAAC
CCGTTCACCCACGGCAGCGCCAAGCACACCGACGTGATGAAGACCGAGGG
ACTCAAGCAGGCCCTGGACAAGTACGGTTTCGACGCTGCCTTCGGCGGTG
CGCGCCGCGACGAGGAGAAGTCGCGGGCCAAGGAACGGGTCTATTCGTTC
CGCGACAGCAAGCACCGCTGGGACCCGAAGAACCAGCGTCCCGAGCTGTG
GAACATCTACAACGGCAAGGTGAAGAAGGGCGAGTCGATCCGCGTCTTCC
CGCTGTCCAACTGGACCGAGCTGGACATCTGGCAATACATCTACCTGGAA
GGCATCCCGATCGTCCCGCTGTACTTCGCCGCCGAGCGCGAGGTCATCGA
GAAGAATGGCACATTGATCATGATCGACGACGAGCGCATCCTCGAGCATC
TCTCTGACGAAGAGAAAGCCCGCATCGAGAAGCGCATGGTGCGCTTCCGT
ACCCTCGGCTGCTACCCGCTCACCGGCGCGGTCGAGTCCAGCGCCACCAC
GCTGCCGGAAATCATCCAGGAAATGCTCCTGACGCGTACTTCCGAACGCC
AGGGCCGGGTCATCGACCATGACCAGGCCGGTTCGATGGAAGAAAAGAAA
CGTCAGGGCTATTTCTGA
[0034] The VIR2 protein (SEQ ID NO:4) encoded by SEQ ID NO:3 is
presented using the one-letter amino acid code in Table 4B.
TABLE-US-00006 TABLE 4B Encoded VIR2 protein sequence (SEQ ID
NO:4)hz,1/32 MVDKLTHLKQLEAESIHIIREVAAEFDNPVMLYSIGKDSAVMLHLARKAF
FPGKLPFPVMHVDTRWKFQEMYRFRDRMVEEMGLDLITHVNPDGVAQGIN
PFTHGSAKHTDVMKTEGLKQALDKYGFDAAFGGARRDEEKSRAKERVYSF
RDSKHRWDPKNQRPELWNIYNGKVKKGESIRVFPLSNWTELDIWQYIYLE
GIPIVPLYFAAEREVIEKNGTLIMIDDERILEHLSDEEKARIEKRMVRFR
TLGCYPLTGAVESSATTLPEIIQEMLLTRTSERQGRVIDHDQAGSMEEKK RQGYF
[0035] The role of VIR2 in virulence was confirmed using phage to
retransduce this mutation into the wild-type PT894 strain where
attenuated virulence was again observed in the Dictyostelium growth
assay compared to an isogenic bacterial strain.
MUT3
[0036] A Pseudomonas bacterial mutant (MUT3) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding CysQ (PA5175). This gene
encodes the VIR3 nucleic acid (SEQ ID NO:5) shown in Table 5A.
TABLE-US-00007 TABLE 5A VIR3 Nucleotide Sequence (SEQ ID NO:5)
ATGAGGCCGGTGCCTTGGGGCGAATTGGTGGCGCTGGTGCGGCGCGCCGGCGAGGCGATCCTGCCGC
ACTGGCGCGCCGACCTGGTGGTGCGCTCGAAGGCCGACGAATCGCCGGTGACTGCCGCCGACCTGGC
CGCGCACCATATATTGGAGGCGGGATTGCGGGCGCTGGCGCCGGACATTCCGGTGCTTTCCGAAGAG
GATTGCGAGATACCGCTGAGCGAGCGCGGCCACTGGCGGCGCTGGTGGCTGGTGGACCCGCTGGACG
GCACCAAGGAGTTCATCTCCGGTAGCGAGGAGTTCACCGTCAACGTGGCCCTGGTCGAGGATGGCCG
GGTGCTGTTCGGCCTGGTCGGCGTGCCGGTGAGCGGCCGCTGCTACTACGGTGGCGCCGGTCTCGGT
GCCTGGCGCGAGGAGGCCGATGGCCGCGCGCAACCGATCAGTGTGCGCCTGGAGCCCGAGGAGGCCT
TCACCGTGCTGGCCAGCAAGCGCCATGGCAGCCCGGCCCAGGAGCGCCTGCTGGATGGCTTGAGCGA
GCGCTTCGGCGACCTGCGGCGAGCCAGCATCGGCAGTTCGCTGAAGTTCTGCCTGCTGGCCGAGGGC
GCTGCCGACTGCTATCCGCGCCTGACGCCAACCTCGCAATGGGACACGGCCGCCGCCCAGGGTGTGC
TGGAAGGCGCCGGCGGCGAGGTGCTCGACCTGCATGGTGCGCCATTCACCTACGAGCCGCGCGAGGA
TTACCTCAACGGCTCCTTCCTGGCCCTGCCGCGCGCCGCCCAGTGGCGCAGCGAGCTGATCCAACTG
GCGCGCGCGCTGCACTGA
[0037] The VIR3 protein (SEQ ID NO:6) encoded by SEQ ID NO:5 is
presented using, the one-letter amino acid code in Table 5B.
TABLE-US-00008 TABLE 5B Encoded VIR3 protein sequence (SEQ ID NO:6)
MRPVPWGELVALVRRAGEAILPHWRADVVVRDKADESPVTAADLAAHHIL
EAGLRALAPDIPVLSEEDCEIPLSERGHWRRWWLVDPLDGTKEFISGSEE
FTVNVALVEDGRVLFGLVGVPVSGRCYYGGAGLGAWREEADGRAQPISVR
LEPEEAFTVVASKRHGSPAQERLLDGLSERFGDLRRASIGSSLKFCLLAE
GAADCYPRLTPTSQWDTAAAQGVLEGAGGEVLDLHGAPFTYEPREDYLNG
SFLALPRAAEWRSELIQLARALH
MUT4
[0038] A Pseudomonas bacterial mutant (MUT4) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding D-amino acid dehydrogenase,
small subunit (dadA; PA5304). This gene encodes the VIR4 nucleic
acid (SEQ ID NO:7) shown in Table 6A. TABLE-US-00009 TABLE 6A VIR4
Nucleotide Sequence (SEQ ID NO:7)
ATGCGAGTTCTGGTCCTTGGCAGCGGTGTCATCGGTACCGCCAGTGCGTATTACCTGGCCCGTGCCG
GGTTCGAGGTGGTGGTGGTCGACCGTCACCACGGTCCCGCGCTGGAAACCAGCTTCGCCAACGCCGG
CCAGGTGTCTCCCGGCTACGCTTCGCCCTGGGCAGCCCCGGGCATTCCCCTGAAGGCCATGAAGTGG
CTGCTGGAAAAGCACGCGCCGCTGGCCATCAAGCTCACCTCCGATCCCAGCCAGTACGCCTGGATGC
TGCAGATGCTGCGCAACTGCACCGCCGAGCGCTACGCCGTGAACAAGGAGCGCATGGTCCGCCTGTC
CGAGTACAGCCGCGATTGCCTCGACGAACTGCGCGCCGAGACCGGCATCGCCTACGAGGGCCGCACC
CTCGGCACCACCCAACTGTTCCGCACCCAGGCGCAGCTGGACGCCGCCGGCAAGGACATCGCCGTGC
TCGAGCGCTCCGGCGTGCCCTACGAGGTTCTCGACCGCGACGGCATCGCCCGCGTAGAGCCGGCTTT
GGCCAAGGTCGCCGACAAGCTGGTCGGCGCCTTGCGCCTGCCCAACGACCAGACCGGCGACTGCCAG
CTGTTCACCACCCGCCTGGCGGAAATGGCCAAGGGCCTGGGCGTGGAGTTCCGCTTCGGCCAGAACA
TCGAGCGCCTGGACTTCGCCGGCGACCGCATCAACGGCGTGCTGGTCAACGGCGAATTGCTCACCGC
CGACCACTACGTGCTGGCCCTGGGCAGCTACTCGCCGCAACTGCTCAAGCCGCTGGGTATCAAGGCT
CCGGTCTATCCGCTGAAGGGTTATTCGCTGACCGTGCCGATCACCAACCCGGAGATGGCGCCGACCT
CGACCATCCTCGACGAGACCTACAAGGTGGCGATCACCCGCTTCGACCAGCGCATCCGCGTCGGCGG
CATGGCGGAAATCGCCGGCTTCGACCTGTCGCTGAACCCGCGCCGCCGCGAGACCCTGGAAATGATC
ACCACCGACCTCTATCCCGAGGGCGGCGATATCAGCCAGGCGACCTTCTGGACCGGCCTGCGCCCGG
CGACCCCGGATGGCACCCCGATCGTCGGCGCCACCCGCTACCGCAACCTGTTCCTCAATACCGGCCA
CGGCACCCTGGGTTGGACCATGGCCTGCGGGTCGGGTCGCTACCTGGCCGACCTGATGGCGAAGAAG
CGCCCGCAGATCAGTACCGAAGGCCTGGATATTTCCCGCTACAGCAATTCCCCGGAGAACGCCAAGA
ATGCCCATCCAGCGCCAGCACACTAA
[0039] The VIR4 protein (SEQ ID NO:8) encoded by SEQ ID NO:7 is
presented using the one-letter amino acid code in Table 6B.
TABLE-US-00010 TABLE 6B Encoded VIR4 protein sequence (SEQ ID NO:8)
MRVLVLGSGVIGTASAYYLARAGFEVVVVDRQDGPALETSFANAGQVSPG
YASPWAAPGIPLKAMKWLLEKHAPLAIKLTSDPSQYAWMLQMLRNCTART
YAVNKERMVRLSEYSRDCLDELRAETGIAYEGRTLGTTQLFRTQAQLDAA
GKDIAVLERSGVPYEVLDRDGIARVEPALAKVADKLVGALRLPNDQTGDC
QLFTTRLAEMAKGLGVEFRFGQNIERLDFAGDRINGVLVNGELLTADHYV
LALGSYSPQLLKPLGIKAPYPLKGYSLTVPITNPEEMAPTSTILDETYKV
AITRFDQRIRVGGMAEIAGFDLSLNPRRRETLEMITTDLYPEGGDISQAT
FWTGLRPATPDGTPIVGATRYRNLFLNTGHGTLGWTMACGSGRYLADLMA
KKRPQISTEGLDISRYSNSPENAKNAHPAPAH
[0040] The role of VIR4 in virulence was confirmed using phage to
retransduce this mutation into the wild-type PT894 strain where
attenuated virulence was again observed in the Dictyostelium growth
assay compared to an isogenic bacterial strain.
MUT5
[0041] A Pseudomonas bacterial mutant (MUT5) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding imidazoleglycerol-phosphate
synthase, cyclase subunit (hisF; PA5140). This gene encodes the
VIR5 nucleic acid (SEQ ID NO:9) shown in Table 7A. TABLE-US-00011
TABLE 7A VIR5 Nucleotide Sequence (SEQ ID NO:9)
ATGGCACTGGCAAAACGCATCATCCCCTGCCTCGACGTGGACAACGGCCGAGTGGTCAAGGGCGTCA
AGTTCGAGAACATCCGCGACGCCGGCGACCCGGTCGAGATCGCTCGCCGCTACGACGAGCAGGGTGC
CGACGAGATCACCTTCCTCGATATCACCGCCAGCGTCGACGGGCGCGACACCACCCTGCATACCGTC
GAGCGCATGGCTAGCCAGGTGTTCATTCCGCTGACCGTGGGCGGCGGCGTACGCAGCGTGCAGGACA
TCCGCAACCTGTTGAATGCCGGCGCGGACAAGGTCTCGATCAACACCGCCGCGGTGTTCAACCCCGA
GTTCGTCGGTGAGGCCGCCGACCGCTTCGGCTCGCAGTGCATCGTGGTCGCCATCGACGCGAAGAAG
GTTTCCGCCCCGGGCGAGGCGCCGCGCTGGGAAATCTTCACCCATGGCGGGCGCAAGCCCACCGGGC
TGGATGCCGTGCTCTGGGCGAAGAAGATGGAAGACTTGGGCGCTGGCGAGATTCTCCTGACCAGCAT
GGACCAGGACGGCGTGAAGAGCGGTTACGACCTGGGCGTGACCCGCGCCATCAGCGAGGCGGTGAAC
GTGCCGGTGATCGCTTCCGGCGGCGTCGGCAACCTGGAGCACCTGGCCGCCGGCATCCTCGAGGGCA
AGGCCGACGCGGTGCTCGCGGCGAGCATCTTCCACTTCGGCGAGTACACCGTGCCGGAAGCCAAGGC
CTACCTGGCCAGCCGCGGTATCGTGGTGCGCTGA
[0042] The VIR5 protein (SEQ ID NO:10) encoded by SEQ ID NO:9 is
presented using the one-letter amino acid code in Table 7B.
TABLE-US-00012 !Encoded VIR5 protein sequence (SEQ ID NO:10)
MALAKRIIPCLDVDNGRVVKGVKFENIRDAGDPVEIARRYDEQGADEITF
LDITASVDGRDTTLHTVERMASQVFIPLTVGGGVRSVQDIRNLLNAGADK
VSINTAAVFNPEFVGEAADRFGSQCIVVAIDAKKVSAPGEAPRWEIFTHG
GRKPTGLDAVLWAKKMEDLGAGEILLTSMDQDGVKSGYDLGVTRAISEAV
NVPVIASGGVGNLEHLAAGILEGKADAVLAASIFHFGEYTVPEAKAYLAS RGIVVR
MUT6
[0043] A Pseudomonas bacterial mutant (MUT6) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding
N-acetyl-.cndot.-glutamyl-phosphate reductase (ArgC; PA0662). This
gene encodes the VIR6 nucleic acid (SEQ ID NO:11) shown in Table
8A. TABLE-US-00013 TABLE 8A VIR6 Nucleotide Sequence (SEQ ID NO:
11)
ATGATCAAGGTCGGCATCGTTGGCGGTACGGGTTATACGGGCGTGGAACTGCTGCGCCTGCTGGCGC
AGCATCCGCAGGCCCGGGTGGAAGTGATCACTTCGCGTTCCGAGGCGGGGGTGAAGGTCGCCGACAT
GTACCCGAACCTGCGAGGTCATTATGACGACCTGCAGTTCAGCGTGCCGGACGCGCAGCGCCTCGGC
GCCTGCCACGTGGTGTTCTTCGCCACGCCGCACGGCGTGGCGCACGCGCTGGCTGGCGAACTGCTGG
ACGCCGGGACCCGGGTCATCGATCTGTCCGCTGACTTCCGCCTGGCGGACGCCGAGGAGTGGGCGCG
CTGGTACGGCCAGCCGCATGGCGCTCCGGCGCTGCTCGACGAGGCTGTCTACGGCCTGCCGGAAGTG
AACCGCGAGAAGATCCGCCAGGCCCGCCTGATCGCCGTGCCGGGCTGCTACCCGACCGCGACCCAGC
TGGGCCTGATCCCGCTGCTCGAAGCCGGCCTGGCCGACGCCTCGCGGCTGATCGCCGATTGCAAGTC
CGGGGTCAGCGGTGCCGGTCGGGGCGCCAAGGTTGGCTCGCTGTTCTGCGAGGCGGGCGAAAGCATG
ATGGCCTACGCGGTCAAAGGGCATCGGCATCTCCCGGAAATCAGCCAGGGCCTGCGTCGGGCCTCCG
GCGGCGACGTCGGGCTGACGTTCGTACCGCACCTGACGCCAATGATCCGCGGTATCCATGCAACCCT
CTATGCCCATGTCGCGGATCGCTCGGTCGACCTCCAGGCGTTGTTCGAGAAGCGCTACGCCGACGAA
CCCTTCGTCGACGTGATGCCGGCCGGCAGCCATCCGGAGACCCGCAGCGTGCGTGGCGCCAATGTCT
GCCGAATCGCCGTGCATCGCCCCCAGGGCGGCGACCTGGTGGTGCTGCTGTCGGTGATCGACAACCT
GGTCAAGGGCGCCTCGGGTCAGGCGCTCCAGAACATGAACATCCTGTTCGGGCTGGACGAGCGCCTG
GGCCTCTCGCATGCGGCCCTGCTCCCCTGA
[0044] The VIR6 protein (SEQ ID NO:12) encoded by SEQ ID NO:11 is
presented using the one-letter amino acid code in Table 8B.
TABLE-US-00014 TABLE 8B Encoded VIR6 protein sequence (SEQ ID NO:
12)
MIKVGIVGGTGYTGVELLRLLAQHPQARVEVITSRSEAGVKVADMYPNLRGHYDDLQFSVPDAQR
LGACDVVFFATPHGVAHALAGELLDAGTRVIDLSADFRLADAEEWARWYGQPHGAPALLDEAVYG
LPEVNREKIRQARLIAVPGCYPTATQLGLIPLLEAGLADASRLIADCKSGVSGAGRGAKVGSLFC
EAGESMMAYAVKGHRHLPEISQGLRRASGGDVGLTFVPHLTPMIRGIHATLYAHVADRSVDLQAL
FEKRYADEPFVDVMPAGSHPETRSVRGANVCRIAVHRPQGGDLVVVLSVIDNLVKGASGQALQNM
NILFGLDERLGLSHAALLP
[0045] The role of VIR6 in virulence was confirmed using phage to
retransduce this mutation into the wild-type PT894 strain where
attenuated virulence was again observed in the Dictyostelium growth
assay compared to an isogenic bacterial strain.
MUT7
[0046] A Pseudomonas bacterial mutant (MM) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding dihydrolipoamide
acetyltransferase (AceF; PA5016). This gene encodes the VIR7
nucleic acid (SEQ ID NO:13) is shown in Table 9A. TABLE-US-00015
TABLE 9A VIR7 Nucleotide Sequence (SEQ ID NO: 13)
GTGAGCGAACTCATTCGCGTACCCGACATCGGCAACGGTGAGGGTGAAGTCATCGAGCTGCTGGTCA
AGCCCGGCGACAAGGTCGAGGCCGATCAGAGCCTGCTGACCCTGGAATCCGACAAGGCCAGCATGGA
AATCCCCAGTCCCAAGGCCGGGGTAGTGAAAAGCATCAAGGCGAAGGTCGGCGACACCTTGAAAGAA
GGTGACGAAATCCTCGAGCTGGAAGTGGAAGGCGGCGAACAGCCTGCCGAAGCCAAGGCCGAGGCAG
CGCCCGCCCAACCGGAAGCGCCGAAAGCCGAAGCGCCTGCTCCCGCCCCGAGCGAGAGCAAGCCGGC
CGCCCCCGCCGCGGCCAGCGTCCAGGACATCAAGGTCCCGGACATCGGCTCGGCCGGCAAGGCCAAC
GTCATCGAAGTGATGGTCAAGGCCGGCGACACGGTCGAGGCCGACCAGTCGCTGATCACCCTGGAAT
CCGACAAGGCCAGCATGGAGATCCCCTCGCCGGCCTCCGGGGTGGTGGAAAGCGTCTCGATCAAGGT
CGGTGACGAAGTCGGCACCGGCGACCTGATCCTCAAGCTGAAGGTGGAAGGCGCCGCTCCGGCAGCC
GAAGAGCAACCGGCAGCCGCTCCGGCCCAGGCCGCGGCGCCCGCCGCCGAGCAGAAGCCCGCCGCGG
CGGCCCCTGCGCCAGCCAAGGCCGATACCCCGGCTCCGGTCGGCGCACCCAGCCGCGACGGCGCCAA
GGTCCACGCCGGCCCGGCGGTGCGCATGCTGGCGCGCGAGTTCGGCGTCGAGCTGAGCGAAGTGAAA
GCCAGCGGTCCCAAGGGTCGCATCCTCAAGGAAGACGTCCAGGTCTTCGTCAAGGAGCAACTGCAGC
GCGCCAAGTCCGGCGGTGCCGGCGCCACCGGCGGAGCCGGCATCCCGCCGATCCCGGAAGTCGACTT
CAGCAAGTTCGGCGAAGTGGAAGAAGTGGCGATGACCCGCCTGATGCAGGTCGGCGCCGCCAACCTG
CATCGCAGCTGGCTGAACGTGCCGCACGTGACCCAGTTCGACCAGTCGGACATCACCGACATGGAAG
CCTTCCGCGTTGCCCAGAAGGCCGCGGCGGAGAAGGCCGGGGTCAAGCTGACCGTACTGCCGATCCT
GCTCAAGGCCTGCGCCCACCTGCTCAAGGAACTGCCGGACTTCAACAGTTCGCTGGCCCCCAGCGGC
AAGGCGCTGATCCGCAAGAAGTACGTACACATCGGCTTCGCCGTGGACACTCCGGACGGCCTGCTGG
TCCCGGTGATCCGCGATGTCGACCGGAAGAGCCTCCTGCAACTGGCCGCCGAGGCCGCCGACCTGGC
CGACAAGGCCCGCAACAAGAAGCTCTCGGCCGATGCCATGCAGGGCGCCTGCTTCACCATCTCCAGT
CTCGGCCACATCGGCGGCACCGGCTTCACGCCGATCGTCAACGCGCCGGAAGTGGCGATCCTCGGTG
TGTCCAAGGCGACCATGCAGCCGGTATGGGACGGCAAGGCCTTCCAGCCGCGCCTGATGCTGCCGCT
GTCGCTGTCCTACGACCATCGCGTGATCAACGGTGCCGCCGCGGCGCGCTTCACCAAGCGCCTGGGC
GAGCTGCTGGCGGACATCCGCACCCTGCTCCTGTAA
[0047] The VIR7 protein (SEQ ID NO:14) encoded by SEQ ID NO:13 is
presented using the one-letter amino acid code in Table 9B.
TABLE-US-00016 TABLE 9B Encoded VIR7 protein sequence (SEQ ID NO:
14)
MSELIRVPDIGNGEGEVIELLVKPGDKVEADQSLLTLESDKASMEIPSPKAGVVKSIKAKVGDTL
KEGDEILELEVEGGEQPAEAKAEAAPAQPEAPKAEAPAPAPSESKPAAPAAASVQDIKVPDIGSA
GKANVIEVMVKAGDTVEADQSLITLESDKASMEIPSPASGVVESVSIKVGDEVGTGDLILKLKVE
GAAPAAEEQPAAAPAQAAAPAAEQKPAAAAPAPAKADTPAPVGAPSRDGAKVHAGPAVRMLAREF
GVELSEVKASGPKGRILKEDVQVFVKEQLQRAKSGGAGATGGAGIPPIPEVDFSKFGEVEEVAMT
RLMQVGAANLHRSWLNVPHVTQFDQSDITDMEAFRVAQKAAAEKAGVLKTVLPILLKACAHLLKE
LPDFNSSLAPSGKALIRKKYVHIGFAVDTPDGLLVPVIRDVDRKSLLQLAAEAADLADKARNKKL
SADAMQGACFTISSLGHIGGTGFTPIVNAPEVAILGVSKATMQPVWDGKAFQPRLMLPLSLSYDH
RVINGAAAARFTKRLGELLADIRTLLL
MUT8
[0048] A Pseudomonas bacterial mutant (MUT8) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding NADH dehydrogenase I chain H
(nuoH; PA2643). This gene encodes the VIR8 nucleic acid (SEQ ID
NO:15) shown in Table 10A. TABLE-US-00017 TABLE 10A VIR8 Nucleotide
Sequence (SEQ ID NO: 15)
ATGAGTTGGCTGACTCCCGCTCTGGTCACCATCATCCTCACCGTGGTCAAGGCCATCGTGGTGCTGC
TCGCCGTGGTCATCTGCGGCGCCCTGCTAAGCTGGGTCGAGCGCCGCCTGCTCGGCCTCTGGCAGGA
CCGCTACGGCCCCAACCGGGTCGGTCCGTTCGGTGCGTTCCAGCTCGGCGCGGACATGGTCAAGATG
TTCTTCAAGGAGGACTGGACCCCGCCGTTCGCCGACAAGATGATCTTCACCCTGGCCCCGGTAATCG
CGATGGGCGCCCTGCTCGTCGCCTTCGCCATCGTGCCGATCACCCCCACCTGGGGCGTGGCGGACCT
GAACATCGGCATCCTGTTCTTCTTCGCCATGGCCGGCCTGACGGTGTACGCCGTGCTGTTCGCCGGC
TGGTCGAGCAACAACAAGTTCGCCCTGCTCGGCAGCCTGCGCGCCTCGGCCCAGACCATCTCCTACG
AGGTGTTCCTGGCCCTGTCGCTGATGGGCATCGTCGCCCAGGTCGGCTCGTTCAACATGCGCGACAT
CGTCCAGTACCAGATCGACAACGTCTGGTTCATCATTCCGCAGTTCTTCGGCTTCTGCACCTTCATC
ATCGCCGGCGTCGCCGTGACCCACCGTCACCCGTTCGACCAGCCGGAAGCGGAGCAGGAACTGGCGG
ACGGCTACCACATCGAGTACGCCGGGATGAAATGGGGCATGTTCTTCGTCGGCGAGTACATCGGCAT
CGTACTGGTCTCGGCGCTGCTGGCGACCCTGTTCTTCGGCGGCTGGCACGGTCCGTTCCTGGACACC
CTGCCCTGGCTGTCGTTCTTCTACTTCGCCGCCAAGACCGGCTTCTTCATCATGCTCTTCATCCTGA
TCCGCGCCTCGCTGCCGCGTCCGCGCTATGACCAGGTGATGGCGTTCAGCTGGAAGGTGTGCCTGCC
GCTGACCCTGATCAACCTGCTGGTGACCGGCGCGCTCGTGCTGGCCGCGGCCCAGTAA
[0049] The VIR8 protein (SEQ ID NO:16) encoded by SEQ ID NO:15 is
presented using the one-letter amino acid code in Table 10B.
TABLE-US-00018 TABLE 10B Encoded VIR8 protein sequence (SEQ ID NO:
16)
MSWLTPALVTIILTVVKAIVVLLAVVICGALLSWVERRLLGLWQDRYGPNRVGPFGAFQLGADMV
KMFFKEDWTPPFADKMIFTLAPVIAMGALLVAFAIVPITPTWGVADLNIGILFFFAMAGLTVYAN
LFAGWSSNNKFALLGSLRASAQTISYEVFLALSLMGIVAQVGSFNMRDIVQYQIDNVWFIIPQFF
GFCTFIIAGVAVTHRHPFDQPEAEQELADGYHIEYAGMKWGMFFVGEYIGIVLVSALLATLFFGG
WHGPFLDTLPWLSFFYFAAKTGFFIMLFILIRASLPRPRYDQVMAFSWKVCLPLTLINLLVTGAL
VLAAAQ
MUT9
[0050] A Pseudomonas bacterial mutant (MUT9) was made by transposon
insertion in a P. aeruginosa wild-type strain PT894. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding pyoverdine synthase D (PvdD;
PA2399). This gene encodes the VIR9 nucleic acid (SEQ ID NO:17)
shown in Table 11A. TABLE-US-00019 TABLE 11A VIR9 Nucleotide
Sequence (SEQ ID NO: 17)
GTGCAAGCACTCATAGAGAAGGTGGGCTCCCTTTCCCCCCAGGAAAGGAAGGCATTGGCTGTCCTGC
TCAAGCAGCAAGGTGTCAATCTCTTCGAGATCGCGCCAGTGTTCAAGCGCCAGGACGGCGAGCCCCT
GCGGCTCTCCTATGCCCAGGAGCGACAGTGGTTTCTCTGGCAACTGGAGCCGGAAAGCGCGGCCTAC
CATATCCCGAGTGTCTTGCGTCTACGTGGGCGGCTGGACCTGGATGCCCTGCAACGCAGCTTCGACA
GCCTGGTTGCGCGGCACGACACCCTACGCACCCGTTTTCGCCTCGACGGCGACGAGGCGCGCCAGGA
GATCGCCGCATCCATGGCATTGCCGTTGGATATCGTCGCGTTGGGGCCGCTCGAGGAGGGCGCCCTC
GCTCGGCAGGTCGAGACGACGATCGCGCGGCCGTTCGACCTGGAGCGTGGGCCGCTGCTGCGGGTGA
GCCTGTTGCGGCTGGCCGAGGACGACCATGTGCTGGTGCTGGTCCAGCATCACATCGTGTCCGACGG
TTGGTCGATGCAGGTGATGGTCGAGGAACTGGTCCAGCTCTATGCCGCCTATAGTCGAGGGCTCGAG
GTAGCGCTGCCGGCTTTGCCGATCCAGTACGCGGACTACGCCCTGTGGCAGCGCAGCTGGATGGAGG
CCGGGGAAAAGGAGCGCCAGTTGGCGTACTGGACCGGCCTGCTGGGCGGCGAGCAGCCGGTGCTGGA
GTTGCCGTTCGACCGGCCCCGCCCGGTTCGGCAAAGCCATCGTGGTGCCCAGTTCATCCTGGAACTG
GATATTGATCTGTCCCAGGCGCTCAGGCGCGTGGCCCAGCAGGAGGGGGCTACTGCCTTCGCCCTGT
TGCTGGCTTCGTTCCAGGCGCTGCTGTATCGCTACAGCGGGCAGGCGGATATCCGTGTCGGCGTGCC
GATCGCCAATCGCAACCGCGTGGAGACCGAGCGGCTGATCGGCTTCTTCGTCAACACCCAGGTGCTC
AAGGCCGACCTGGACGGTCGGATGGGCTTCGACGAGCTGCTGGCCCAGGCCCGCCAACGCGCGCTGG
AGGCCCAGGCGCACCAGGACCTGCCGTTCGAGCAACTGGTGGAGGCCTTGCAGCCGGAGCGCAGTCT
TAGCCACAACCCGCTGTTCCAGGTGCTGTTCAACTACCAGAGCGAAGCCCGTGGCAACGGCCAGGCA
TTCCGCTTCGACGAGTTACAGATGGAAAGCGTGCAGTTCGACAGCCGGACGGCGCAGTTCGACTTGA
CGTTGGACCTGACGGACGAAGAGCAGCGTTTTTGCGCCGTTTTCGACTACGCCACCGACCTGTTCGA
CGCCTCCACCGTGGAACGCCTGGCCGGCCATTGGCGCAACCTGTTGCGCGGCATCGTCGCCAACCCA
CGACAGCGGCTCGGCGAGTTGCCGCTGCTGGATGCGCCGGAGCGCCGGCAGACCCTCTCCGAATGGA
ACCCGGCCCAGCGCGAGTGCGCGGTGCAGGGCACCTTGCAGCAGCGTTTCGAGGAACAGGCGCGGCA
ACGGCCACAGGCGGTTGCGCTGATCCTCGACGAACAACGGTTGAGCTACGGCGAACTGAATGCGCGG
GCCAATCGCCTGGCGCACTGCCTGATCGCCCGTGGCGTTGGCGCGGACGTGCCGGTCGGGCTGGCGC
TGGAGCGTTCGCTGGACATGCTGGTCGGCTTGCTGGCGATCCTCAAGGCCGGCGGCGCCTACCTGCC
GTTGGACCCGGCGGCGCCAGAGGAGCGCCTGGCGCATATCCTCGACGACAGTGGGGTACGGCTGCTG
CTGACCCAGGGGCATCTGCTCGAGCGCCTGCCACGGCAGGCGGGGGTGGAGGTGCTGGCCATCGACG
GACTGGTGCTGGACGGCTACGCCGAGAGCGATCCGCTCCCGACGCTATCGGCGGACAACCTGGCCTA
CGTGATCTATACCTCGGGCTCGACCGGCAAGCCCAAGGGCACATTGCTCACCCACCGCAACGCGCTG
CGCCTGTTCAGCGCCACCGAGGCCTGGTTCGGCTTCGACGAGCGGGACGTGTGGACATTGTTCCATT
CCTACGCCTTCGATTTCTCGGTCTGGGAAATCTTCGGCGCGCTGCTCTATGGCGGGTGCCTGGTGAT
TGTGCCGCAATGGGTGAGCCGTTCGCCGGAAGACTTCTACCGTCTGCTGTGCCGCGAAGGCGTGACG
GTGCTCAACCAGACGCCGTCGGCGTTCAAGCAACTGATGGCGGTGGCCTGTTCCGCCGACATGGCGA
CGCAGCAGCCGGCGCTGCGCTACGTGATCTTCGGTGGCGAGGCGCTGGATCTGCAGAGCCTGCGGCC
GTGGTTCCAGCGCTTCGGCGATCGCCAGCCGCAACTGGTGAACATGTACGGCATCACCGAGACCACG
GTGCACGTAACCTACCGTCCGGTGAGCGAGGCCGACCTGGAAGGTGGCCTGGTCAGTCCGATTGGCG
GGACCATCCCGGACCTGTCCTGGTACATCCTCGACCGTGACCTGAACCCGGTGCCGCGCGGCGCGGT
GGGCGAGCTGTACATCGGTCGCGCCGGGCTGGCGCGCGGCTACCTGAGGCGGCCCGGGTTGAGTGCC
ACCCGCTTCGTGCCGAACCCGTTCCCCGGCGGCGCCGGCGAGCGGCTGTACCGTACCGGCGACCTGG
CACGGTTCCAGGCGGATGGCAATATCGAGTACATCGGGCGTATCGACCACCAGGTGAAGGTTCGCGG
CTTCCGTATCGAACTGGGCGAGATCGAAGCGGCGCTCGCCGGTCTGGCCGGGGTACGCGATGCCGTG
GTGCTGGCCCATGACGGAGTCGGCGGCACGCAACTGGTGGGATACGTGGTGGCGGACTCGGCGGAGG
ATGCCGAGCGTCTGCGGGAGTCGCTGCGGGAGTCGCTGAAGCGGCACCTGCCGGACTACATGGTGCC
GGCGCACCTGATGCTGCTGGAGCGGATGCCGCTGACGGTCAATGGCAAGCTCGACCGGCAGGCGTTG
CCGCAACCGGATGCGAGCCTGTCGCAACAGGCCTATCGAGCGCCCGGTAGCGAGCTGGAGCAGCGCA
TCGCAGCGATCTGGTCGGAGATCCTGGGAGTGGAACGGGTCGGCCTGGACGACAACTTCTTCGAACT
GGGCGGTCATTCGTTGCTGGCTACCCGGGTGATTTCTCGGGTTCGCCAGGAGCAGCAGTTGGACGCA
AGCCTGAAGGCGTTGTTCGAGCGGCCGGTTCTGGAAGCGTTCGCCCAGGGATTGGAACGCACGACGG
ATGCGGTCTCGACGATACCGCTTGCCGATCGGCAGCAACCGTTGGCACTGTCCTTCGCTCAGGAGCG
TCAGTGGTTCCTCTGGCAACTGGAGCCGGAAAGCGCGGCCTACCATATTCCGAGTGCCTTGCGCCTA
CGCGGGCGGCTGGACGTGGATGCCTTGCAACGCAGCTTCGACAGCCTGGTCGCGCGGCATGAAACCT
TGCGTACCCGCTTCCGGCTGGAGGGAGGGCGTTCGTACCAGCAGGTACAACCTGCGGTTAGCGTTTC
CATCGAGCGGGAACAGTTCGGTGAAGAAGGCCTGATCGAACGGATACAGGCCATCGTTGTGCAGCCA
TTCGACCTGGAACGGGGGCCGCTGCTGCGGGTGAACCTGTTGCAACTGGCCGAGGACGACCATGTAC
TGGTGCTGGTCCAGCACCACATCGTGTCCGATGGTTGGTCGATGCAGGTGATGGTCGAGGAACTGGT
CCAGCTCTATGCCGCCTATAGCCAAGGGCTCGACGTGGTGTTGCCAGCCCTGCCGATCCAGTACGCG
GACTACGCCCTGTGGCAGCGCAGCTGGATGGAGGCGGGGGAAAAGGAGCGCCAGTTGGCGTACTGGA
CCGGCCTGCTGGGCGGCGAGCAGCCGGTGCTGGAGTTGCCGTTCGATCGGCCGCGTCCGGCCCGGCA
GAGCCATCGTGGCGCGCAGTTGGGTTTCGAGCTATCGCGGGAACTGGTCGAGGCCGTGAGAGCCTTG
GCCCAGCGTGAAGGCGCCAGTAGTTTCATGCTGTTGCTGGCCTCGTTCCAGGCGCTGTTGTATCGCT
ACAGCGGGCAGGCGGATATCCGTGTCGGTGTGCCGATCGCCAATCGCAACCGCGTGGAGACCGAGCG
GCTGATCGGCTTCTTCGTCAACACCCAGGTGCTCAAGGCCGACCTGGACGGTCGGATGGGCTTCGAC
GAGCTGCTGGCCCAGGCCCGCCAACGCGCGCTGGAGGCCCAGGCGCACCAGGACCTGCCGTTCGAGC
AACTGGTGGAAGCCTTGCAGCCGGAGCGCAATGCCAGCCACAACCCACTGTTCCAGGTGCTGTTCAA
CCATCAGAGCGAGATACGCTCGGTGACGCCCGAGGTTCAGTTGGAGGACCTGCGTCTGGAAGGCCTG
GCCTGGGACGGCCAGACTGCGCAGTTCGACCTGACGCTGGATATTCAGGAAGACGAAAACGGCATCT
GGGCCTCCTTCGACTATGCCACCGATCTGTTCGACGCCTCCACCGTGGAACGCCTGGCCGGCCATTG
GCGCAACCTGTTGCGCGGCATCGTCGCCAACCCACGACAGCGGCTCGGCGAGTTGCCGCTGCTGGAT
GCGCCGGAGCGCCGGCAGACCCTCTCCGAATGGAACCCGGCCCAGCGCGAGTGCGCGGTGCAGGGCA
CCTTGCAGCAGCGTTTCGAGGAGCAGGCGCGGCAACGGCCACAGGCGGTTGCGCTGATCCTCGACGA
ACAACGGTTGAGCTACGGCGAACTGAATGCGCGGGCCAATCGCCTGGCGCACTGCCTGATCGCTCGC
GGCGTTGGCGCGGACGTGCCGGTCGGGCTGGCGCTGGAGCGTTCGCTGGACATGCTGGTCGGCTTGC
TGGCGATCCTCAAGGCCGGCGGCGCCTACCTGCCGTTGGACCCGGCGGCGCCACAGGAGCGCCTGGC
GCATATCCTCGACGACAGTGGGGTACGGCTGCTGCTGACCCAGGGGCATCTGCTCGAGCGCCTGCCG
CGGCAGGCGGGGGTGGAGGTGCTGGCCATCGACGGACTGGTGCTGGACGGCTACGCCGAGAGCGATC
CGCTCCCGACGCTATCGGCGGACAACCTGGCCTACGTGATCTATACCTCGGGCTCGACCGGCAAGCC
CAAGGGCACGTTGCTCACCCACCGCAACGCGCTGCGCCTGTTCAGCGCCACCGAGGCCTGGTTCGGC
TTCGACGAGCGGGACGTGTGGACGTTGTTCCATTCCTACGCCTTCGATTTCTCGGTCTGGGAAATCT
TCGGCGCGCTGCTCTATGGCGGGCGCCTGGTGATCGTGCCGCAATGGGTGAGCCGTTCGCCGGAAGA
CTTCTACCGTCTGCTGTGCCGCGAAGGCGTGACGGTGCTCAACCAGACGCCGTCGGCGTTCAAGCAA
CTGATGGCGGTGGCCTGTTCCGCCGACATGGCGACGCAGCAGCCGGCGCTGCGCTACGTGATCTTCG
GTGGCGAGGCGCTGGATCTGCAGAGCCTGCGGCCGTGGTTCCAGCGCTTTGGCGATCGCCAGCCGCA
ACTGGTGAACATGTACGGCATCACCGAGACCACGGTACACGTAACCTACCGTCCGGTGAGCGAAGCC
GACCTGAAGGGTGGCCTGGTCAGTCCGATCGGCGGGACCATCCCGGACCTGTCCTGGTACATCCTCG
ACCGTGACCTGAACCCGGTGCCGCGCGGCGCGGTGGGCGAGCTGTACATCGGTCGCGCCGGTCTGGC
GCGCGGCTACCTGAGGCGGCCCGGGTTGAGTGCCACCCGCTTCGTGCCGAACCCGTTCCCCGGCGGT
GCCGGCGAGCGGCTGTACCGTACCGGCGACCTGGCACGGTTCCAGGCGGATGGCAATATCGAGTACA
TCGGGCGTATCGACCACCAGGTGAAGGTTCGCGGCTTCCGTATCGAACTGGGTGAGATCGAAGCGGC
GCTCGCCGGTCTGGCCGGGGTACGCGATGCCGTGGTGCTGGCCCATGACGGGGTCGGCGGCACGCAA
CTGGTGGGATACGTGGTGGCGGACTCGGCGGAGGATGCCGAGCGTCTGCGGGAGTCGCTGCGGGAGT
CGCTGAAGCGGCACCTGCCGGACTACATGGTGCCGGCGCACCTGATGCTGCTGGAGCGGATGCCGCT
GACGGTCAATGGCAAGCTCGACCGGCAGGCGTTGCCGCAACCGGATGCGAGCTTGTCGCAGCAGGCC
TATCGAGCGCCCGGTAGCGAGCTGGAGCAGCGCATCGCAGCGATCTGGGCGGAGATCCTGGGAGTGG
AACGGGTCGGCCTGGACGACAACTTCTTCGAACTGGGCGGTCACTCATTGTTGCTGCTGATGCTCAA
GGAGCGGATCGGCGATACCTGCCAGGCTACGCTGAGCATCAGCCAACTGATGACCCATGCCAGCGTC
GCGGAACAGGCGGCATGCATCGAGGGGCAGGCGCGTGAGTCGTTGCTGGTGCCGCTCAACGGCAGGC
GCGAAGGTTCGCCGCTGTTCATGTTCCATCCGAGTTTCGGCTCTGTGCACTGTTACAAGACCCTCGC
CATGGCGCTGCGGGATCGTCATCCGGTCAAGGGTGTTGTCTGCCGTGCCCTGCTGGGCGCTGGTCGC
GAGGTGCCGGAGTGGGACGATATGGTTGCGGAATACGCCGAGCAATTGCTGCAGGAGCACCCCGAAG
GGGTTTTCAACCTGGCGGGATGGTCGCTCGGCGGCAACCTGGCGATGGATGTCGCGGCCCGGCTGGA
GCAGCGTGGGCGGCAGGTGGCTTTCGTCGGCTGGATCGATGCACCGGCACCGGTCAGGGTCGAAGCG
TTCTGGAACGAGATCGGGCCGACGCCGGAGGCAGTCCCGAACCTATCCGTGGGCGAGATGCGGGTGG
AACTGCTCGGTGTCATGTTTCCGGAGCGGGCCGAGCATATCGAACGGGCCTGGTCATCGATCTGCTC
CGCCACGACGGACGATGAGCAGCGCTGGACGAGGATGAGCGACTGGGCGGAAGCGGAGATCGGCGCC
GAGTTCGCGACACTGCGCAGCGAAATCGCACAGAGCAACGAACTGGAAGTGTCCTGGGAGTTGAAAC
AGATCCTCGACGAGCGCCTGAAAGCGATGGATTACCCGCGTCTGACGGCGAAGGTCAGCCTCTGGTG
GGCCGCGCGCAGCACCAATGCCATCCAGCGGAGCGCGGTGGAGCGCTCGATGGCCGAGGCGATCGGG
GCTGAGCGTGTCGAACCGGTGCGGGTGCTGGATACCCGGCACGACAAGATCATCGACCACCCTGAGT
TTGTGCAGAGCTTCCGGGCCGCCCTGGAGCGTGCCGGGCGCTGA
[0051] The VIR9 protein (SEQ ID NO:18) encoded by SEQ ID NO:17 is
presented using the one-letter amino acid code in Table 11B.
TABLE-US-00020 TABLE 11B Encoded VIR9 protein sequence (SEQ ID NO:
18)
MQALIEKVGSLSPQERKALAVLLKQQGVNLFEIAPVFKRQDGEPLRLSYAQERQWFLWQLEPESA
AYHIPSVLRLRGRLDLDALQRSFDSLVARHETLRTRFRLDGDEARQEIAASMALPLDIVALGPLE
EGALARQVETTIARPFDLERGPLLRVSLLRLAEDDHVLVLVQHHIVSDGWSMQVMVEELVQLYAA
YSRGLEVALPALPIQYADYALWQRSWMEAGEKERQLAYWTGLLGGEQPVLELPFDRPRPVRQSHR
GAQFILELDIDLSQALRRVAQQEGATAFALLLASFQALLYRYSGQADIRVGVPIANRNRVETERL
IGFFVNTQVLKADLDGRMGFDELLAQARQRALEAQAHQDLPFEQLVEALQPERSLSHNPLFQVLF
NYQSEARGNGQAFRFDELQMESVQFDSRTAQFDLTLDLTDEEQRFCAVFDYATDLFDASTVERLA
GHWRNLLRGIVANPRQRLGELPLLDAPERRQTLSEWNPAQRECAVQGTLQQRFEEQARQRPQAVA
LILDEQRLSYGELNARANRLAHCLIARGVGADVPVGLALERSLDMLVGLLAILKAGGAYLPLDPA
APEERLAHILDDSGVRLLLTQGHLLERLPRQAGVEVLAIDGLVLDGYAESDPLPTLSADNLAYVI
YTSGSTGKPKGTLLTHRNALRLFSATEAWFGFDERDVWTLFHSYAFDFSVWEIFGALLYGGCLVI
VPQWVSRSPEDFYRLLCREGVTVLNQTPSAFKQLMAVACSADMATQQPALRYVIFGGEALDLQSL
RPWFQRFGDRQPQVLNMYGITETTVHVTYRPVSEADLEGGLVSPIGGTIPDLSWYILDRDLNPVP
RGAVGELYIGRAGLARGYLRRPGLSATRFVPNPFPGGAGERLYRTGDLARFQADGNIEYIGRIDH
QVKVRGFRIELGEIEAALAGLAGVRDAVVLAHDGVGGTQLVGYVVADSAEDAERLRESLRESLKR
HLPDYMVPAHLMLLERMPLTVNGKLDRQALPQPDASLSQQAYRAPGSELEQRIAAIWSEILGVER
VGLDDNFFELGGHSLLATRVISRVRQEQQLDASLKALFERPVLEAFAQGLERTTDAVSTIPLADR
QQPLALSFAQERQWFLWQLEPESAAYHIPSALRLRGRLDVDALQRSFDSLVARHETLRTRFRLEG
GRSYQQVQPAVSVSIEREQFGEEGLIERIQAIVVQPFDLERGPLLRVNLLQLAEDDHVLVLVQHH
IVSDGWSMQVMVEELVQLYAAYSQGLDVVLPALPIQYADYALWQRSWMEAGEKERQLAYWTGLLG
GEQPVLELPFDRPRPARQSHRGAQLGFELSRELVEAVRALAQREGASSFMLLLASFQALLYRYSG
QADIRVGVPIANRNRVETERLIGFFVNTQVLKADLDGRMGFDELLAQARQRALEAQAHQDLPFEQ
LVEALQPERNASHNPLFQVLFNHQSEIRSVTPEVQLEDLRLEGLAWDGQTAQFDLTLDIQEDENG
IWASFDYATDLFDASTVERLAGHWRNLLRGIVANPRQRLGELPLLDAPERRQTLSEWNPAQRECA
VQGTLQQRFEEQARQRPQAVALILDEQRLSYGELNARANRLAHCLIARGVGADVPVGLALERSLD
MLVGLLAILKAGGAYLPLDPAAPEERLAHILDDSGVRLLLTQGHLLERLPRQAGVEVLAIDGLVL
DGYAESDPLPTLSADNLAYVIYTSGSTGKPKGTLLTHRNALRLFSATEAWFGFDERDVWTLFHSY
AFDFSVWEIFGALLYGGRLVIVPQWVSRSPEDFYRLLCREGVTVLNQTPSAFKQLMAVACSADMA
TQQPALRYVIFGGEALDLQSLRPWFQRFGDRQPQLVNMYGITETTVHVTYRPVSEADLKGGLVSP
IGGTIPDLSWYILDRDLNPVPRGAVGELYIGRAGLARGYLRRPGLSATRFVPNPFPGGAGERLYR
TGDLARFQADGNIEYIGRIDHQVKVRGFRIELGEIEAALAGLAGVRDAVVLAHDGVGGTQLVGYV
VADSAEDAERLRESLRESLKRHLPDYMVPAHLMLLERMPLTVNGKLDRQALPQPDASLSQQAYRA
PGSELEQRIAAIWAEILGVERVGLDDNFFELGGHSLLLLMLKERIGDTCQATLSISQLMTHASVA
EQAACIEGQARESLLVPLNGRREGSPLFMFHPSFGSVHCYKTLAMALRDRHPVKGVVCRALLGAG
REVPEWDDMVAEYAEQLLQEHPEGVFNLAGWSLGGNLAMDVAARLEQRGRQVAFVGWIDAPAPVR
VEAFWNEIGPTPEAVPNLSVGEMRVELLGVMFPERAEHIERAWSSICSATTDDEQRWTRMSDWAE
AEIGAEFATLRSEIAQSNELEVSWELKQILDERLKAMDYPRLTAKVSLWWAARSTNAIQRSAVER
SMAEAIGAERVEPVRVLDTRHDKIIDHPEFVQSFRAALERAGR
[0052] A Pseudomonas bacterial mutant (MUT10) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding the RND multidrug efflux
transporter MexD (mexD; PA4598). This gene encodes the VIR10
nucleic acid (SEQ ID NO:19) shown in Table 12A. TABLE-US-00021
TABLE 12A VIR10 Nucleotide Sequence (SEQ ID NO: 19)
ATGTCCGAATTCTTCATCAAGCGGCCGAACTTCGCCTGGGTGGTGGCCCTGTTCATCTCCCTGGCCG
GCCTGCTGGTCATTTCCAAATTGCCGGTAGCGCAGTACCCCAATGTCGCGCCGCCACAGATCACCAT
CACCGCCACCTATCCCGGCGCCTCGGCGAAGGTGCTGGTGGACTCCGTCACCAGTGTGCTCGAGGAG
TCGCTGAACGGCGCCAAGGGCCTGCTCTACTTCGAGTCGACCAACAACTCCAACGGCACCGCCGAGA
TCGTCGTCACCTTCGAGCCGGGCACCGATCCGGACCTGGCCCAGGTGGACGTGCAGAACCGCCTGAA
GAAAGCCGAGGCGCGCATGCCGCAGGCGGTGCTGACCCAGGGCCTGCAGGTCGAGCAGACCAGCGCC
GGTTTCCTGCTGATCTATGCGCTCAGCTACAAGGAAGGCGCTCAGCGCAGCGACACCACCGCCCTCG
GCGACTACGCCGCGCGCAATATCAACAACGAGCTGCGGCGCCTGCCGGGCGTCGGCAAGCTGCAATT
CTTCTCTTCCGAGGCGGCCATGCGGGTCTGGATCGATCCGCAGAAGCTGGTGGGCTTCGGCCTCTCC
ATCGACGACGTGAGCAATGCCATCCGCGGGCAGAACGTGCAGGTGCCGGCCGGCGCCTTCGGCAGCG
CACCGGGCAGTTCCGCGCAGGAGCTGACGGCGACCCTGGCGGTGAAGGGCACCCTGGACGATCCGCA
GGAGTTCGGCCAGGTAGTGCTGCGCGCCAACGAGGACGGCTCGCTGGTCCGGCTCGCCGATGTCGCG
CGCCTGGAACTCGGCAAGGAGAGCTACAACATTTCCTCGCGACTGAACGGCACGCCCACCGTGGGCG
GGGCTATCCAGCTGTCGCCCGGGGCCAACGCGATCCAGACCGCTACCCTGGTGAAACAGCGTCTCGC
CGAACTGTCGGCGTTCTTCCCCGAGGACATGCAGTACAGCGTGCCCTACGACACCTCGCGCTTCGTC
GACGTGGCCATCGAGAAGGTGATCCACACCCTGATCGAAGCGATGGTCCTGGTGTTCCTGGTGATGT
TCCTGTTCCTGCAGAACGTCCGCTACACCCTGATCCCGTCCATCGTGGTGCCGGTGTGCCTGCTGGG
TACGCTGATGGTGATGTACCTGCTGGGGTTCTCGGTGAACATGATGACCATGTTCGGCATGGTCCTG
GCGATCGGCATCCTGGTGGACGACGCCATCGTGGTGGTGGAGAACGTCGAGCGGATCATGGCGGAGG
AGGGGATTTCCCCGGCCGAGGCCACGGTCAAGGCGATGAAGCAGGTATCCGGCGCCATCGTCGGCAT
CACCCTGGTGCTCTCGGCGGTGTTCCTGCCGCTGGCTTTCATGGCCGGTTCGGTGGGGGTGATCTAC
CAGCAGTTCTCGGTGTCGCTGGCGGTCTCGATCCTGTTCTCCGGCTTCCTCGCCCTGACCTTCACCC
CGGCGCTGTGCGCCACGCTGCTCAAGCCCATTCCCGAAGGGCACCACGAGAAGCGCGGCTTCTTCGG
CGCCTTCAACCGTGGCTTCGCCCGCGTCACCGAGCGCTATTCGCTGCTCAACTCGAAGCTGGTGGCG
CGCGCCGGACGCTTCATGCTGGTGTACGCCGGCCTGGTGGCCATGCTCGGCTACTTCTACCTGCGCC
TGCCGGAAGCCTTCGTGCCGGCGGAAGACCTCGGCTACATGGTGGTCGACGTGCAACTGCCGCCTGG
CGCTTCGCGCGTGCGCACCGATGCCACCGGCGAGGAGCTCGAGCGCTTCCTCAAGTCCCGCGAGGCG
GTGGCTTCGGTGTTCCTGATCTCGGCTTCAGCTTCTCCGGCCAGGGCGACAATGCCGCCGCTGGCCT
TCCCAACCTTCAAGGACTGGTCCGAGCGAGGCGCCGAGCAGTCGGCCGCCGCCGAGATCGCCGCGCT
GAACGAGCATTTCGCGCTGCCCGACGATGGCACGGTCATGGCCGTGTCGCCGCCACCGATCAACGGT
CTGGGTAACTCCGGCGGCTTCGCATTGCGCCTGATGGACCGTAGCGGGGTCGGCCGCGAAGCGCTGC
TGCAGGCTCGCGATACTCTTCTTGGCGAGATCCAGACCAACCCGAAATTCCTTTACGCGATGATGGA
AGGACTGGCCGAAGCGCCGCAACTGCGCCTGTTGATCGACCGGGAGAAGGCCCGTGCCCTGGGGGTG
AGCTTCGAGACCATCAGCGGCACGCTGTCCGCTGCCTTCGGCTCGGAGGTGATCAACGACTTCACCA
ATGCGGGGCGCCAACAGCGGGTGGTGATCCAGGCCGAACAGGGCAACCGGATGACCCCGGAAAGCGT
GCTCGAGCTATACGTGCCTAACGCTGCTGGCAACCTGGTACCGCTCAGCGCCTTCGTCAGCGTGAAA
TGGGAAGAGGGACCGGTGCAATTGGTGCGCTATAACGGCTACCCGTCGATCCGCATCGTCGGTGACG
CCGCGCCCGGCTTCAGTACCGGCGAAGCCATGGCGGAAATGGAGCGCCTGGCCTCGCAGCTGCCGGC
CGGCATCGGCTACGAGTGGACCGGCCTGCTCCTATCAGGAGAAGGTCTCCGCCGGCAGGCCACCAGC
CTGTTCGCCCTCGCCATCCTGGTGGTGTTCCTGTTGCTGGTGGCGCTCTACGAGAGCTGGTCGATCC
CGCTGTCGGTGATGCTGATCGTGCCGATCGGCGCCATCGGCGCGGTGCTCGCGGTGATGGTCAGCGG
TATGTCCAACGACGTGTATTTCAAGGTCGGCCTGATCACCATCATCGGTCTTTCGGCGAAGAACGCG
ATCCTCATCGTCGAGTTCGCCAAGGAACTCTGGGAGCAGGGGCATAGCCTGCGCGACGCCGCCATCG
AGGCCGCGCGCCTGCGCTTCCGGCCGATCATCATGACTTCCATGGCGTTCATCCTCGGCGTGATACC
CCTGGCCCTGGCCAGCGGTGCCGGCGCGGCGAGCCAGCGTGCCATCGGCACCGGAGTGATCGGCGGG
ATGCTCAGCGCCACCTTCCTCGGCGTGCTGTTCGTACCTATCTGTTTCGTCTGGCTGCTGTCGCTGC
TGCGCAGCAAGCCGGCACCCATCGAACAGGCCGCTTCGGCCGGGGAGTGA
[0053] The VIR10 protein (SEQ ID NO:20) encoded by SEQ ID NO:19 is
presented using the one-letter amino acid code in Table 12B.
TABLE-US-00022 TABLE 12B Encoded VIR10 protein sequence (SEQ ID NO:
20)
MSEFFIKRPNFAWVVALFISLAGLLVISKLPVAQYPNVAPPQITITATYPGASAKVLVDSVTSVL
EESLNGAKGLLYFESTNNSNGTAEIVVTFEPGTDPDLAQVDVQNRLKKAEARMPQAVLTQGLQVE
QTSAGFLLIYALSYKEGAQRSDTTALGDYAARNINNELRRLPGVGKLQFFSSEAAMRVWIDPQKL
VGFGLSIDDVSNAIRGQNVQVPAGAFGSAPGSSAQELTATLAVKGTLDDPQEFGQVVLRANEDGS
LVRLADVARLELGKESYNISSRLNGTPTVGGAIQLSPGANAIQTATLVKQRLAELSAFFPEDMQY
SVPYDTSRFVDVAIEKVIHTLIEAMVLVFLVMFLFLQNVRYTLIPSIVVPVCLLGTLMVMYLLGF
SVNMMTMFGMVLAIGILVDDAIVVVENVERIMAEEGISPAEATVKAMKQVSGAIVGITLVLSAVF
LPLAFMAGSVGVIYQQFSVSLAVSILFSGFLALTFTPALCATLLKPIPEGHHEKRGFFGAFNRGF
ARVTERYSLLNSKLVARAGRFMLVYAGLVAMLGYFYLRLPEAFVPAEDLGYMVVDVQLPPGASRV
RTDATGEELERFLKSREAVASVFLISGFSFSGQGDNAALAFPTFKDWSERGAEQSAAAEIAALNE
HFALPDDGTVMAVSPPPINGLGNSGGFALRLMDRSGVGREALLQARDTLLGEIQTNPKFLYAMME
GLAEAPQLRLLIDREKARALGVSFETISGTLSAAFGSEVINDFTNAGRQQRVVIQAEQGNRMTPE
SVLELYVPNAAGNLVPLSAFVSVKWEEGPVQLVRYNGYPSIRIVGDAAPGFSTGEAMEAMERLAS
QLPAGIGYEWTGLSYQEKVSAGQATSLFALAILVVFLLLVALYESWSIPLSVMLIVPIGAIGAVL
AVMVSGMSNDVYFKVGLITIIGLSAKNAILIVEFAKELWEQGHSLRDAAIEAARLRFRPIIMTSM
AFILGVIPLALASGAGAASQRAIGTGVIGGMLSATFLGVLFVPICFVWLLSLLRSKPAPIEQAAS
AGE
MUT11
[0054] A Pseudomonas bacterial mutant (MUT11) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding PA3721. This gene encodes the
VIR11 nucleic acid (SEQ ID NO:21) shown in Table 13A.
TABLE-US-00023 TABLE 13A VIR11 Nucleotide Sequence (SEQ ID NO: 21)
ATGAACGATGCTTCTCCCCGTCTGACCGAACGCGGCAGGCAACGCCGCCGCGCCATGCTCGACGCCG
CTACCCAGGCCTTTCTCGAACACGGTTTCGAAGGCACCACCCTGGACATGGTGATAGAACGGGCCGG
TGGTTCACGGGGGACCCTGTACAGCTCCTTCGGCGGCAAGGAGGGCCTGTTCGCCGCGGTGATCGCC
CACATGATCGGGGAAATCTTCGACGACAGCGCCGATCAGCCGCGCCCCGCCGCCACGCTGAGCGCCA
CCCTCGAGCATTTCGGCCGGCGCTTTCTCACCAGCCTGCTCGATCCCCGCTGCCAGAGCCTCTATCG
CCTGGTGGTGGCGGAATCCCCGCGGTTTCCGGCGATCGGCAAGTCCTTCTACGAGCAGGGGCCGCAG
CAGAGCTATCTGCTGCTCAGCGAGCGACTGGCCGCGGTCGCTCCTCACATGGACGAGGAAACGCTCT
ACGCGGTGGCCTGCCAGTTTCTCGAGATGCTCAAGGCCGACCTGTTCCTCAAGGCCCTCAGCGTGGC
CGACTTCCAGCCGACCATGGCGCTGCTGGAAACCCGCCTCAAGCTGTCGGTGGACATCATCGCCTGC
TACCTGGAACACCTGTCGCAGAGCCCCGCGCAGGGCTGA
[0055] The VIR11 protein (SEQ ID NO:22) encoded by SEQ ID NO:21 is
presented using the one-letter amino acid code in Table 13B.
TABLE-US-00024 TABLE 13B Encoded VIR11 protein sequence (SEQ ID NO:
22)
MNDASPRLTERGRQRRRAMLDAATQAFLEHGFEGTTLDMVIERAGGSRGTLYSSFGGKEGLFAAV
IAHMIGEIFDDSADQPRPAATLSATLEHFGRRFLTSLLDPRCQSLYRLVVAESPRFPAIGKSFYE
QGPQQSYLLLSERLAAVAPHMDEETLYAVACQFLEMLKADLFLKALSVADFQPTMALLETRLKLS
VDIIACYLEHLSQSPAQG
MUT12
[0056] A Pseudomonas bacterial mutant (MUT12) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding PA0596. This gene encodes the
VIR12 nucleic acid (SEQ ID NO:23) shown in Table 14A.
TABLE-US-00025 TABLE 14A VIR12 Nucleotide Sequence (SEQ ID NO: 23)
ATGTCTGATGATGCCCGTTTCCAGCAGCTGAATTGCTGGTTGGACTCTTGTTTGCCCGAGTTGTTCG
TTGCCGAAGGTTGGGGGGAAGTGCCCCCCGCCGAACTGATCCCGGCCAGTAGCGACGCCAGCTTCCG
TCGTTATTTCCGCTGGCAGGGAGGGGACCGCAGCCTGGTGGTGATGGACGCGCCGCCGCCCCAGGAA
GACTGCCGACCGTTCGTCAAGGTCGCCGGACTGCTCGCCGGAGCCGGCGTGCATGTGCCGAGGATTC
TCGCCCAGGACCTGGAGAACGGTTTCCTGCTGCTCAGTGACCTGGGCCGGCAGACCTACCTCGACGT
GCTTCATCCCGGGAATGCCGACGAGCTGTTCGAACCGGCCCTGGATGCGCTGATCGCCTTCCAGAAG
GTCGATGTCGCCGGTGTCCTGCCTGCCTACGACGAAGCGGTGCTGCGCCGCGAGCTGCAGCTGTTCC
CCGACTGGTACCTGGCCCGCCACCTCGGCGTGGAGCTGGAGGGCGAGACGCTGGCCCGCTGGAAACG
GATCTGCGACCTGCTGGTACGCAGCGCGCTGGAGCAACCGCGGGTGTTCGTCCATCGCGACTATATG
CCGCGCAATCTGATGCTCAGCGAGCCCAACCCGGGCGTCCTCGACTTCCAGGACGCCCTGCACGGCC
CGGTCACCTACGATGTCACCTGCCTGTACAAGGACGCCTTCGTCAGTTGGCCGGAGCCGCGCGTGCA
TGCCGCGCTGAACCGTTACTGGAAGAAGGCGACCTGGGCCGGCATCCCGCTGCCGCCAAGCTTCGAA
GACTTCCTCCGTGCCAGCGACCTGATGGGCGTGCAGCGCCACCTGAAGGTGATTGGCATCTTCGCCC
GTATCTGTCACCGCGACGGCAAGCCGCGCTACCTGGGTGACGTGCCGCGCTTCTTCCGTTATCTGGA
AACCGCCGTGGCGCGCCGTCCCGAGCTGGCCGAACTGGGCGAGCTGCTGGCCTCGCTGCCGCAGGGA
GCCGAGGCATGA
[0057] The VIR12 protein (SEQ ID NO:24) encoded by SEQ ID NO:23 is
presented using the one-letter amino acid code in Table 14B.
TABLE-US-00026 TABLE 14B Encoded VIR12 protein sequence (SEQ ID NO:
24)
MSDDARFQQLNCWLDSCLPELFVAEGWGEVPPAELIPASSDASFRRYFRWQGGDRSLVVMDAPPP
QEDCRPFVKVAGLLAGAGVHVPRILAQDLENGFLLLSDLGRQTYLDVLHPGNADELFEPALDALI
AFQKVDVAGVLPAYDEAVLRRELQLFPDWYLARHLGVELEGETLARWKRICDLLVRSALEQPRVF
VHRDYMPRNLMLSEPNPGVLDFQDALHGPVTYDVTCLYKDAFVSWPEPRVHAALNRYWKKATWAG
IPLPPSFEDFLRASDLMGVQRHLKVIGIFARICHRDGKPRYLGDVPRFFRYLETAVARRPELAEL
GELLASLPQGAEA
MUT13
[0058] A Pseudomonas bacterial mutant (MUT13) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding PA5265. This gene encodes the
VIR13 nucleic acid (SEQ ID NO:25) shown in Table 15A.
TABLE-US-00027 TABLE 15A VIR13 Nucleotide Sequence (SEQ ID NO: 25)
ATGAGCGGATTCCAGGACCAGAGTATCGACGAAGGCGTGCGCAAGCGCACCGCCTACCAGAACGATC
GGCGTGCACGACTGGCATTGAACGTCGAGCGACAGGACGGCGGTATCCTGCAGATTCCGGTGGCCAG
CGATATGCTCGGCCATGAGGAGCACGAGCGTATCCAGCAGAACACCTTCCTGGCTGTGATGCCGCTG
GTCCGCCTGCCAACGCTGGGCAAGGCCGGTTATGGCGACCAGCTGCCCGCCGGCGCGCTACCGCGGG
CGGGACGGATCTACCTGTTCCAGGACGGCAAGTTGTGGCGCGAACTGGAATGTGATGGCAAGGGCAA
CCTGTTCGAAGTCGATCTCCTGCAGGGGCGCAGCCAGCGTGCGGACAAGCGTCCGGCCTTAGGCAAG
ACACAAGCGCTGATCCTGGTGCCGGTGCTGGTCAAGGGGCAGTTCGTGATCCCACGCTACACCATGG
CCTATAGCGAAACTCCCTGGCCTTGGTCGTACATCGACTGGCTGGAGGAGGACCCGCAGCGGGTCAA
CCGGCGCTGCCAGCAGATGGCGTCCGCTTGGAACGCCTCGGTGGCCAACCAGCACTGGAAAGCCTCC
ATCCATCAACCCGCGCTGGTCATTGATCATCACGCCCAGGGTTTGCGACCTCGCGACTTCAACGTCG
AGAGCGCGCTGGAAGACCCGGCGGAATTCACACCTGAGTTCGCCGCCTTTCGCGAAGAGTCGCTGGT
GTGCCAGTTGCAGCGACGCCAGCAGGAATTGGCGCCCCTGCTGAAGCAGGCTCCGCCCTCTGCGCTA
CCTACTCTGGAAGCCGGAGAGGACGTACTGGAAACCCTCAAGCTGCGTGGCCATCCCAACCTCATCG
GGCTGATGCTCGACGACTCGCTGTTCGCCTTGCGCCACGCTGCGGCGCAGGCGCGCCACTGCGCCGC
CTACTTGCGCAGCCTCAATGCACTGCTGCCGCACCGTCCCAACGGACGCTATGCACAGGTGCTGAGC
AACATGCTCGACGGCCCGCTCGCCAAGCTCAGGGGCGAGGTCGATCAGGCCGAACTGGACGAGGCGA
TCTTCGCCGAGGAGCGACAGTCTTGCCGAATCCACCTGACGCAGCAGGTCGAGCATCTGGTTGCCCT
GCTGGAAGGCCCCTTGCACCCGGTGTTGCAGGACTGGACCCACCAGTGCGACGAAGCCCTGCTGGAG
CCCTACAGCCTGATGAGCGAGGCACTGGCTGCGCTGAACCAGCTTCCCGACCGCTGCGACGCACTGT
ACAGCGGTACCGCCTACCGGGCGCTGGCGGCACATGTCGAGCGGGTGGTCAGCACGGTTCTGCAGGC
AAGCCACCCGCTTGGCGCCATGCTCCTGGCCAAGGACGAAGGACAACTTCCCGAGCCGGTTCGGCGC
CTGCAGGCGCTGCGCGATAGCCCGCGGACGCCGGACCCCGATGCAATGGGCCTCAGCACGCTGATGC
TGGGAGCCAGTCTGCTGGGCGAGGTCGACCAGCCCAGCGCCGGCAAGAGCCTCGCCTACTTCCTCGG
CGACCTGCTGGACGTGTTCGGCGCCAGCGTAGTCGAGCAACTCGGCCGGCTGTCCCAGGGCGCCACC
CAGATCCAGCTCGACCGCTTGTTCGCACCGACCTTCAATACTCTGAGCGCCCTCTCGGTGAAGATGA
AAGGTATCCGCCTGCTGCCCGACAGTCAGGTGCCGCTCGACATGGTTGTCGTCGGCGTGCGCGGAGC
CGGCCTGCGCAACGGTCTGACCGAGGTCGAGCGCCAGGAGCTGAGGCGCAAGAGCTATCGGCGCGCC
ATCGTTCAGGACGGTGCCGGCAATCCCCTGGCCGGCACCAGTCCCCGCGACACCGGCATGAGTCGCG
CCAACCTGCGCAACGTCATGGTGGTGGCGGTACCCAAGGATCACCCGGACCTGCTTGCCTACACGAA
ATTCCGTACGCAGTTAGGCACGTTGACCCAGGTGATGGAGAACACTCGCATCGTGCCGACGATGATG
CTGGGGTTTGCGATTTATAACTTGAATGTGCAGGTGCAGGCATACAGTGGCTTTGTAGACAGTGGAG
AAAAGCACAGAGGGACGATCGGGGCTGTCGGTGCAGTAATCGATTTAACAGCCGCTGGAGGAAGCCA
TGCAAAGCTGCTTTTCGGACCATCTACTGCAAAGTATCTAGAAACCCCACGTATATCGGTAGCCCAA
ATATCCCCTCGATGGGCCAGGAATCTAGAAGTTCAAACAGGCAGCCCTAAGTTAGGGTTGCTACGTG
GGCTTGGTGGCGCAGCCACACTATTCGGTGCAGGCATCAGTGTATGGGATGGCTACCGAGCTTTGAG
GCAGGGAGATAGCGATGCGGCTGCGGCCTACGGTGTGGCCGCAGTGGGTGGGGGCCTTTGGGGTGCC
TACGTCCTAGGATGGATAGTAAACCCTTATGCTTTGCTGGCTGGTGCGGTTTTGGCGATCGGAGGCA
CTGTGGTCGCTAATCTACTGACTGACAGCGATGCGGAAACCATCGTAAAGAAAGGCCCCTTCGGCCG
GCAATTCGCCGAGGCTGGCCTGCTCGATTCGCTGATGGGCCAGGACCAGCGCTTCGCCCATCTGAAA
GACCCGCAAACGGCCTATCGCCAATTGCTGGGAGTCCTCGGCCATCCGCGGGTCTTTGTCCATCGCC
TGGAAGACTGGCGCAAATTGGCGCCGGCGGCGCATCGATCTGTCTTGCAGGAAGCGGAACGGGGTCG
CCAAGCGGTCAGCCGCACTGCGCTATCCTGCATCGACCCCAAGTTGCAGGCGCTGGAGGCAAACGAT
TGGGCCGTGGTGCTGAGTTCCCCGCTCCTGGCCATGTTCGAGAATGGCCAGAAGGCGTTCCGCCTGG
TGGCCCAGGAGTTTCTCAGCAGCTTGCCGATCGATCCGGGCACCCTGTTCGGCGTCAAGCGCTACCA
TCGGGTCCCCGCGGGCCCCGCCAAGCTCGAAGCCTTGCCGTTGGATGCTGCCAGCGTGCTCTATGTG
CTGCCGGCCAGCCTGCCGATTCCGCAGTTGTCTCCTCGGGCCCGCTATAGCATGCGCATGACCCAGG
GTTTGAAGATCAGCGCACAGTTCGAACTCAATGCCGACCAGCCTGAGCAGCGGCTTGTCCTGCCTCA
ACCCAGCCCGAAGAGTTGGAGTGCATTCACATCCGCCAATCGGTACCTTCCCCCGGACGACTTGGGC
CCCCATGCTGCGCCACCTTATTGGTTGATAGAGAACAGTGAGTTCAACGTATGA
[0059] The VIR13 protein (SEQ ID NO:26) encoded by SEQ ID NO:25 is
presented using the one-letter amino acid code in Table 15B.
TABLE-US-00028 TABLE 15B Encoded VIR13 protein sequence (SEQ ID NO:
26)
MSGFQDQSIDEGVRKRTAYQNDRRARLALNVERQDGGILQIPVASDMLGHEEHERIQQNTFLAVM
PLVRLPTLGKAGYGDQLPAGALPRAGRIYLFQDGLKWRELECDGKGNLFEVDLLQGRSQRADKRP
ALGKTQALILVPVLVKGQFVIPRYTMAYSETPWPWSYIDWLEEDPQRVNRRCQQMASAWNASVAN
QHWKASIHQPALVIDHHAQGLRPRDFNVESALEDPAEFTPEFAAFREESLVCQLQRRQQELAPLL
KQAPPSALPTLEAGEDVLETLKLRGHPNLIGLMLDDSLPALRHAAAQARHCAAYLRSLNALLPHR
PNGRYAQVLSNMLDGPLAKLRGEVDQAELDEAIFAEERQSCRIHLTQQVEHLVALLEGPLHPVLQ
DWTHQCDEALLEPYSLMSEALAALNQLPDRCDALYSGTAYRALAAHVERVVSTVLQASHPLGAML
LAKDEGQLPEPVRRLQALRDSPRTPDPDAMGLSTLMLGASLLGEVDQPSAGKSLAYFLGDLLDVF
GASVVEQLGRLSQGATQIQLDRLFAPTFNTLSALSVKMKGIRLLPDSQVPLDMVVVGVRGAGLRN
GLTEVERQELRRKSYRRAIVQDGAGNPLAGTSPRDTGMSRANLRNVMVVAVPKDHPDLLAYTKFR
TQLGTLTQVMENTRIVPTMMLGFAIYNLNVQVQAYSGFVDSGEKHRGTIGAVGAVIDLTAAGGSH
AKLLFGPSTAKYLETPRISVAQISPRWARNLEVQTGSPKLGGLRGLGGAATLFGAGISVWDGYRA
LRQGDSDAAAAYGVAAVGGGLWGAYVLGWIVNPYALLAGAVLAIGGTVVANLLTDSDAETIVKKG
PFGRQFAEAGLLDSLMGQDQRFAHLKDPQTAYRQLLGVLGHPRVFVHRLEDWRKLAPAAHRSVLQ
EAERGRQAVSRTALSCIDPKLQALEANDWAVVLSSPLLAMFENGQKAFRLVAQEFLSSLPIDPGT
LFGVKRYHRVPAGPAKLEALPLDAASVLYVLPASLPIPQLSPRARYSMRMTQGLKISAQFELNAD
QPEQRLVLPQPSPKSWSAFTSANRYLPPDDLGPHAAPPYWLIENSEFNV
MUT14
[0060] A Pseudomonas bacterial mutant (MUT14) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding pyochelin biosynthetic protein
pchC (PA4229). This gene encodes the VIR14 nucleic acid (SEQ ID
NO:27) shown in Table 16A. TABLE-US-00029 TABLE 16A VIR14
Nucleotide Sequence (SEQ ID NO: 27)
ATGAGCGCCGCCTGGGTCCGGCCGTTCCGCCTGACGCCGATGCCGCGCCTGCGCCTGGCCTGCTTCC
CCCATGCAGGCGGCAGCGCCAGCTTCTTCCGTAGCTGGAGCGAACGCCTGCCGCCAGACATCGACCT
GCTTGCCCTGCAGTACCCGGGTCGCGAGGACCGCTTCAACGAGGCGCCGGCCACCCGCCTGGAGGAC
CTCGCCGACGGCGCCGCCCTCGCCCTGCGCGATTTCGCCGACGCGCCCCTGGCGCTGTTCGGCCACA
GTCTCGGCGCGGCGCTGGCCTACGAAACCGCCCTGCGCCTGGAAAGCGCCGGCGCGCCGCTGCGCCA
CCTGTTCGTCTCCGCCCATCCGGCACCGCACCGGCAACGCGGCGGCGCGTTGCACCGCGGCGACGAG
GCGGCGCTGCTGGAGGACGTCCGCCGCCAGGGTGGCGCCAGCGAGCTACTCGAGGACGCCGACCTGC
GCGCGCTGTTCCTGCCGATCCTGCGCGCCGACTACCAGGCGATCGAGACCTACCGACGGGCGCAGCC
CATCGCCCTGGCCTGCGCCCTCGACGTCCTCCTCGGCGAGCACGACGAGGAAGTCAGCGCCGCCGAG
GCGCAGGCCTGGAGCGACGCCAGCCGGACTCCCGCCAGGCTGCGGCGCTTTCCTGGCGGCCACTTCT
ACCTGAGCGAGGGGCGCGACGCGGTGATCGAGCACCTGCTGCGCCGCCTCGCACATCCCGACGCCCT
TTCCCGAGAGGTTGCATGA
[0061] The VIR14 protein (SEQ ID NO:28) encoded by SEQ ID NO:27 is
presented using the one-letter amino acid code in Table 16B.
TABLE-US-00030 TABLE 16B Encoded VIR14 protein sequence (SEQ ID NO:
28)
MSAAWVRPFRLTPMPRLRLACFPHAGGSASFFRSWSERLPPDIDLLALQYPGREDRFNEAPATRLEDL
ADGAALALRDFADAPLALFGHSLGAALAYETALRLESAGAPLRHLFVSAHPAPHRQRGGALHRGDEAA
LLEDVRRQGGASELLEDADLRALFLPILRADYQAIETYRRAQPIALACALDVLLGEHDEEVSAAEAQA
WSDASRTPARLRRFPGGHFYLSEGRDAVIEHLLRRLAHPDALSREVA
MUT15
[0062] A Pseudomonas bacterial mutant (MUT15) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding dihydroaeruginoic acid
synthetase pchE (PA4226). This gene encodes the VIR15 nucleic acid
(SEQ ID NO:29) shown in Table 17A. TABLE-US-00031 TABLE 17A VIR15
Nucleotide Sequence (SEQ ID NO: 29)
ATGGATCTGCCCCCCGATTCCCGTACCGCCCTGCGCGACTGGCTGACCGAGCAGCTCGCCGACCTGC
TCGGCGAACCGCTTGCTGACGTGCGCGCCCTGGCGGACGACGACGACCTGCTGGGCTGCGGCCTCGA
CTCGATCCGCCTGATGTACCTGCAGGAACGCCTGCGCGCGCGTGGCTCGACGCTGGACTTCGCCCAG
TTGGCGCAGCGCCCCTGCCTGGGGGCCTGGCTCGACCTGCTGGCCTGCGCGGACCGGCTGTCCGCCC
CGGCAACGGTCGCGCTGCCGACGGCGCAGGATCGCGATCAGCCGTTCGAGCTGTCTTCCGTGCAGCA
GGCCTACTGGCTGCGACGTGGCGCCGGCGAGGTGCTGGGCAACGTCAGCTGCCATGCCTTTCTGGAA
TTCCGCACGCGGGATGTCGACCCGCAGCGCCTGGCCGCGGCGGCGGAGTGCGTGCGTCAACGCCACC
CGATGTTGCGGGCGCGCTTCCTCGACGGTCGCCAGCAGATCCTTCCGACGCCGCCGCTGTCCTGCTT
CGACCTGCAGGACTGGCGCACCTTACAGGTGGACGAGGCCGAGCGCGACTGGCAGGCGCTGCGCGAC
TGGCGCGCCCATGAATGCCTGGCGGTGGAGCGCGGCCAGGTGTTCCTGCTCGGGCTGGTGCGCATGC
CGGGCGGCGAGGATCGCCTCTGGCTGAGTCTCGACCTGCTTGCCGCCGATGTCGAAAGCCTCCGCCT
GCTGCTGGCCGAACTGGGCGTTGCCTACCTGGCGCCGGAGCGCCTGGCGGAGCCGCCGGCGCTGCAT
TTCGCCGACTACCTGGCGCACCGTGCGGCGCAACGCGCCGAGGCCGCGGCGCGGGCCCGCGACTACT
GGCTGGAACGCCTGCCGCGCTTGCCGGACGCGCCGGCCCTGCCGTTGGCCTGCGCGCCGGAAAGCAT
CCGCCAGCCGCGCACCCGGCGCCTGGCATTCCAGCTTTCCGCCGGCGAGAGCCGGCGCCTGGAGCGT
CTTGCCGCGCAGCATGGCGTGACCTTGTCCAGCGTGTTCGGCTGCGCCTTCGCGCTGGTCCTGGCGC
GCTGGAGCGAAAGCGCGGAATTTCTCCTCAACGTGCCGTTGTTCGATCGGCATGCCGACGACCCGCG
TATCGGCGAGGTGATCGCCGACTTCACCACCCTGTTGCTGCTGGAGTGCCGGATGCAGGCCGGGGTG
TCCTTCGCCGAGGCGGTGAAGAGCTTCCAGCGCAACCTCCACGGAGCCATCGACCACGCCGCATTCC
CCGCCCTGGAGGTGCTCCGCGAGGCGCGCCGGCAGGGCCAGCCACGCTCGGCGCCGGTGGTGTTCGC
CAGCAACCTGGGCGAGGAGGGCTTCGTCCCGGCGGCCTTCCGCGACGCTTTCGGCGATCTCCACGAC
ATGCTCTCGCAGACCCCGCAGGTCTGGCTCGACCACCAGCTCTACCGGGTGGGCGACGGTATCCTGC
TGGCCTGGGATAGCGTCGTCGGCCTGTTCCCCGAAGGTCTGCCGGAAACCATGTTCGAAGCCTACGT
GGGGCTGCTCCAGCGTCTCTGCGACAGCGCCTGGGGGCAGCCCGCCGATCTGCCGTTGCCCTGGGCG
CAGCAGGCGCGCCGGGCCCTGCTCAACGGCCAGCCGGCATGCGCCACGGCGCGCACCCTGCATCGCG
ACTTCTTCCTTCGCGCCGCCGAGGCGCCGGATGCCGACGCGCTGCTCTATCGCGACCAACGTGTCAC
CCGCGGCGAACTGGCCGAGCGTGCGCTGCGCATCGCCGGCGGCCTGCGCGAAGCCGGGGTGCGCCCT
GGCGACGCGGTCGAGGTCAGCCTGCCGCGCGGACCGCAGCAGGTCGCGGCGGTATTCGGCGTGCTCG
CCGCAGGCGCCTGCTACGTGCCGCTGGACATCGACCAGCCGCCCGCACGGCGGCGCCTGATCGAAGA
GGCCGCCGGGGTATGCCTGGCGATCACCGAGGAGGACGATCCGCAGGCCTTGCCGCCGCGCCTGGAT
GTCCAGCGCCTGCTGCGCGGCCCGGCGCTGGCCGCCCCCGTGCCGCTGGCGCCGCAGGCGAGTGCCT
ATGTGATCTACACCTCGGGCTCCACCGGGGTGCCCAAGGGCGTCGAGGTCAGCCACGCGGCGGCGAT
CAATACCATCGACGCGCTGCTCGACCTGCTGCGGGTGAACGCATCGGATCGCTTGCTGGCGGTCTCC
GCGCTGGACTTCGATCTGTCGGTCTTCGACCTGTTCGGCGGCCTCGGCGCCGGTGCCAGCCTGGTCC
TGCCGGCCCAGGAACAGGCGCGCGATGCCGCTGCCTGGGCGGAGGCTATCCAGCGGCATGCGGTGAG
CCTGTGGAACTCGGCGCCGGCCTTGCTGGAGATGGCCCTCAGCCTGCCGGCGAGCCAGGCCGACTAT
CGCAGTCTGCGGGCGGTGCTGCTGTCCGGCGACTGGGTGGCCCTGGACCTGCCCGGCCGCCTGCGCC
CACGTTGTGCCGAAGGCTGCCGCCTGCATGTGCTGGGTGGCGCTACCGAAGCGGGCATCTGGTCGAA
CCTGCAGAGCGTCGATACGGTGCCGCCGCACTGGCGTTCGATTCCCTACGGCCGGCCATTGCCGGGA
CAGGCCTACCGGGTGGTCGACACCCACGGGCGCGACGTGCCGGACCTGGTGGTCGGCGAGCTGTGGA
TCGGCGGCGCCAGCCTGGCCCGCGGCTATCGCAACGATCCCGAACTCAGCGCCCGGCGTTTCGTCCA
CGATGCCCAGGGCCGCTGGTATCGCACCGGCGATCGCGGTCGCTACTGGGGCGACGGTACCCTGGAA
TTCCTCGGTCGGGTCGACCAGCAGGTGAAAGTGCGCGGCCAGCGCATCGAGTTGGGCGAGGTGGAGG
CCGCGCTGTGCGCCCAGGCTGGCGTGGAGAGCGCCTGCGCGGCGGTGCTCGGCGGTGGCGTGGCGAG
CCTCGGCGCGGTGCTGGTACCGCGCCTGGCGCCACGGGCCGAAGGCTCCATGGATCTACCGGCCGCA
CAGCCCTTCGCCGGCCTGGCAGAGGCCGAGGCGGTACTCACCCGGGAAATCCTCGGCGCGCTGCTGG
AGGCGCCGCTGGAGCTAGACGACGGTTTGCGCCGGCGCTGGCTGGACTGGCTAGCGGACTCCGCCGC
CAGCGCGCTGCCGTCGCTCGACGAGGCGTTGCGCCGGCTCGGCTGGCAGGCCGCGGGGCTGACCGCG
ATGGGCAACGCTCTGCGCGGCCTGCTCGCCGGCGAACAGGCGCCGGCCGCGCTGCTCCTCGATCCCT
GGCTGGCGCCGCAGGCGGTGGCCGCGCGCCTGCCGGACGGCCGCGAGGCCCTGGCGCGCCTGCTCGA
AGCGCTGCCGACGCCGGCTGCCGGCGAACGCCTGCGGGTGGCGGTGCTGGATACCCGCGCCGGGCTC
TGGCTCGACCAGGGCATGGCCTCGCTGTTGCGCCCAGGGCTGGAACTGACCCTCTTCGAACGCAGCC
GCGTCCTCCTCGACGCCGCCGCCACCCGCTTGCCGGAACGGATCGTGGTGCAGGCGCTGGACGACGG
CCTGCTACCTGCCGAGCACCTCGGTCGCTACGACCGGGTGATCAGCTTCGCCGCGCTGCACGCCTAC
GAGGCCAGCCGCGAAGGCCTGGCGCTGGCGGCGGCGCTGCTGCGCCCGCAGGGCCGCCTGTTGCTGG
TGGACCTGCTATGCGAGTCGCCACTGGCGCTGCTCGGTGCGGCCTTGCTCGACGACCGGCCGCTGCG
CCTGGCGGAGCTGCCGAGCCTGTTGGCCGATCTCGCCGCTGCGGGACTGGCGCCGCGTTGCCTGTGG
CGCAGCGAGCGGATCGCCCTGGTCGAGGCGCTGGCACCGGGACTCGGGCTCGACGCCGCCGCGCTCC
AGGCCGGCCTGGAGCAACGCCTGCCCCAGGCGATGCGGCCCGAACGCCTGTGGTGCCTGCCAAGCCT
GCCGTTGAACGGCAATGGCAAGGTCGATCGTCGCCGCCTGGCCGAGAGCATGACCCGCGCACTCGGC
GAGTGTCGTCACGAGCCCTCGGCGGAGGAGCCGCTGGAAGCCCATGAGCAAGCGCTGGCCGAGTGCT
GGGAAGCGGTTCTCAAACGCCCGGTGCGTCGTCGCGAGGCGAGCTTCTTCAGCCTCGGCGGCGACAG
CCTGCTGGCGACCCGCCTGCTGGCCGGCATACGTGAGCGTTTCGGCGTACGCCTGGGCATGGCCGAC
TTCTATCGCCAGCCGACCCTGGCCGGTCTTGCCCGCCACTTGCAGGTGCAGACCGTCGAAATCGAGG
AAACCCAACTGGAAGAGGGCGTGCTATGA
[0063] The VIR15 protein (SEQ ID NO:30) encoded by SEQ ID NO:29 is
presented using the one-letter amino acid code in Table 17B.
TABLE-US-00032 TABLE 17B Encoded VIR15 protein sequence (SEQ ID NO:
30)
MDLPPDSRTALRDWLTEQLADLLGEPLADVRALADDDDLLGCGLDSIRLMYLQERLRARGSTLDFAQL
AQRPCLGAWLDLLACADRLSAPATVALPTAQDRDQPFELSSVQQAYWLGRGAGEVLGNVSCHAFLEFR
TRDVDPQRLAAAAECVRQRHPMLRARFLDGRQQILPTPPLSCFDLQDWRTLQVDEAERDWQALRDWRA
HECLAVERGQVFLLGLVRMPGGEDRLWLSLDLLAADVESLRLLLAELGVAYLAPERLAEPPALHFADY
LAHRAAQRAEAAARARDYWLERLPRLPDAPALPLACAPESIRQPRTRRLAFQLSAGESRRLERLAAQH
GVTLSSVFGCAFALVLARWSESAEFLLNVPLFDRHADDPRIGEVIADFTTLLLLECRMQAGVSFAEAV
KSFQRNLHGAIDHAAFPALEVLREARRQGQPRSAPVVFASNLGEEGFVPAAFRDAFGDLHDMLSQTPQ
VWLDHQLYRVGDGILLAWDSVVGLFPEGLPETMFEAYVGLLQRLCDSAWGQPADLPLPWAQQARRALL
NGQPACATARTLHRDFFLRAAEAPDADALLYRDQRVTRGELAERALRIAGGLREAGVRPGDAVEVSLP
RGPQQVAAVFGVLAAGACYVPLDIDQPPARRRLIEEAAGVCLAITEEDDPQALPPRLDVQRLLRGPAL
AAPVPLAPQASAYVIYTSGSTGVPKGVEVSHAAAINTIDALLDLLRVNASDRLLAVSALDFDLSVFDL
FGGLGAGASLVLPAQEQARDAAAWAEAIQRHAVSLWNSAPALLEMALSLPASQADYRSLRAVLLSGDW
VALDLPGRLRPRCAEGCRLHVLGGATEAGIWSNLQSVDTVPPHWRSIPYGRPLPGQAYRVVDTHGRDV
PDLVVGELWIGGASLARGYRNDPELSARRFVHDAQGRWYRTGDRGRYWGDGTLEFLGRVDQQVKVRGQ
RIELGEVEAALCAQAGVESACAAVLGGGVASLGAVLVPRLAPRAEGSMDLPAAQPFAGLAEAEAVLTR
EILGALLEAPLELDDGLRRRWLDWLADSAASALPSLDEALRRLGWQAAGLTAMGNALRGLLAGEQAPA
ALLLDPWLAPQAVAARLPDGREALARLLEALPTPAAGERLRVAVLDTRAGLWLDQGMASLLRPGLELT
LFERSRVLLDAAATRLPERIVVQALDDGLLPAEHLGRYDRVISFAALHAYEASREGLALAAALLRPQG
RLLLVDLLCESPLALLGAALLDDRPLRLAELPSLLADLAAAGLAPRCLWRSERIALVEALAPGLGLDA
AALQAGLEQRLPQAMRPERLWCLPSLPLNGNGKVDRRRLAESMTRALGECRHEPSAEEPLEAHEQALA
ECWEAVLKRPVRRREASFFSLGGDSLLATRLLAGIRERFGVRLGMADFYRQPTLAGLARHLQVQTVEI
EETQLEEGVL
MUT16
[0064] A Pseudomonas bacterial mutant (MUT16) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding pyochelin synthetase pchF
(PA4225). This gene encodes the VIR16 nucleic acid (SEQ ID NO:31)
shown in Table 18A. TABLE-US-00033 TABLE 18A VIR16 Nucleotide
Sequence (SEQ ID NO: 31)
ATGAGCCTCGGCGAACTGCTGGAAACCTGCCGCAGCCGGCGCATCGAACTCTGGAGCGAGGCGGGCC
GCCTGCGCTATCGCGCCCCCCAGGGTGCCCTCGACGCCGGCCTCGCCGAGCGCCTGCGGGCCGAGCG
CGAGGCCCTGCTGGAACACCTGGAAGGCGGCCCTGGCTGGCGCGCCGAACCCGACATGGCCCACCAG
CGCTTCCCGCTGACCCCGGTGCAGGCCGCCTACGTGCTGGGCCGCCAGGCGGCCTTCGACTACGGCG
GTAACGCCTGCCAGCTGTACGCCGAGTACGACTGGCCGGCCGACACCGATCCGGCGCGCCTGGAGGC
GGCCTGGAACGCCATGGTCGAGCGCCACCCGATGCTGCGCGCGGTGATCGAGGACAACGCCTGGCAG
CGCGTGCTGCCCGAGGTGCCCTGGCAGCGGCTGACCGTGCATGCCTGCGCGGGGCTCGACGAGGCCG
CTTTCCAGGCGCACCTGGAGCGGGTCCGCGAACGCCTCGACCACGCCTGCGCGGCGCTCGACCAGTG
GCCGGTCCTGCGCCCCGAGCTGAGTATCGGCCGGGATGCCTGCGTACTGCACTGCTCGGTGGATTTC
ACCCTGGTCGACTACGCCAGCCTGCAATTGCTGCTTGGCGAATGGCGCCGCCGCTATCTCGATCCGC
AATGGACGGCGGAACCGCTGGAGGCGACCTTCCGCGACTATGTCGGCGTCGAGCAGCGCCGACGCCA
GTCGCCAGCCTGGCAGCGCGACCGCGACTGGTGGCTGGCGCGTCTCGACGCGCTACCGGGGCGTCCC
GACCTGCCGCTGCGGGTGCAGCCGGACACCCGGTCCACGCGCTTCCGGCACTTCCACGCGCGCCTCG
ACGAGGCCGCCTGGCAGGCGCTCGGCGCGCGCGCCGGCGAACACGGCCTGAGCGCTGCCGGCGTGGC
CTTGGCGGCCTTCGCCGAGACCATCGGTCGCTGGAGCCAGGCACCGGCGTTCTGTCTCAACCTGACG
GTACTCAACCGGCCGCCGCTGCATCCGCAGCTGGCGCAGGTGCTCGGTGACTTCACCGCGCTCAGCC
TGCTGGCAGTGGACAGCCGCCACGGCGACAGTTTCGTCGAGCGTGCCCGACGCATCGGCGAGCAGAT
GTTCGACGACCTCGACCACCCGACCTTCAGCGGCGTCGACCTGCTGCGCGAACTGGCGCGCCCGCGT
GGTCGCGGCGCCGATCTCATGCCGGTGGTGTTCACCAGTGGCATCGGCAGCGTGCAGCGCCTGCTCG
GCGATGGCGAGGCGCCGCGCGCGCCACGCTACATGATCAGCCAGACCCCGCAGGTCTGGCTGGACTG
CCAGGTCACCGACCAGTTCGGCGGCCTGGAGATCGGCTGGGACGTACGCCTCGGGTTGTTCCCCGAG
GGCCAGGCGGAAGCCATGTTCGACGACTTCGTCCGGCTGCTCCGGCGCCTGGCGCAGAGCCCGCGCG
CCTGGACCGACGGCGATGCCACGGAACCCGTCGAGGCGCCGCCGCAGGCGTTGCCCGGTAGTGCCCG
GAGCATCGCCGCCGGTTTCGCCGAGCGTGCCCTGCTGACCCCCGACGCCACGGCGATCCACGATGCC
GCCGGCAGCTACAGCTACCGCCAGCTCGCCCAGCACGCCAGCGCCCTGCGCCGCGTCCTGGAAGCGC
ACGGCGCGGGCCGTGGCCGGCGGGTCGCGGTGATGCTGCCGAAAAGCGCCGCGCAATTGGTCGCGGT
GATCGGCATCCTCCAGGCCGGCGCCGCCTATGTGCCGGTGGACATCCGCCAGCCTCCGCTGCGGCGC
CAGGCGATCCTCGCCAGCGCCGAAGTGGTCGCGCTGGTCTGCCTGGAAAGCGATGTCCCGGACGTCG
GCTGCGCCTGCGTGGCCATCGACCGGCTGGCCGCCGACAGCGCCTGGCCGCCACCGCCCGCGGCGGA
GGTGGCGGCGGACGACCTCGCCTACGTGATCTACACCTCCGGCTCCACCGGCACGCCAAAGGGCGTG
ATGCTCAGCCATGCGGCGGTGAGCAACACGCTGCTCGACATCAACCAGCGCTACGGCGTCGACGCCA
ACGACCGCGTCCTCGGCCTCGCCGAGCTGAGCTTCGACCTCTCGGTCTACGACTTCTTCGGCGCCAC
CGCGGCGGGGGCCCAGGTGGTCCTCCCGGACCCGGCGCGCGGCAGCGATCCATCGCACTGGGCGGAA
CTGCTGGAACGCCACGCCATCACCCTGTGGAACTCGGTGCCGGCCCAAGGCCAGATGCTCATCGATT
ACCTGGAGAGCGAGCCGCAACGTCACCTGCCGGGACCGCGCTGCGTGCTCTGGTCCGGTGACTGGAT
TCCGGTCAGCCTGCCGACCCGCTGGTGGCGGCGCTGGCCGGACAGCGCGCTGTTCAGCCTGGGCGGC
GCCACCGAGGCGGCGATCTGGTCGATCGAGCAGCCGATCCGCCCGCAGCACACCGAGCTGGCCAGCA
TCCCTTATGGCCGTGCCCTGCGCGGGCAGAGCGTGGAAGTCCTGGATGCCCGCGGGCGGCGCTGCCC
GCCGGGCGTGCGCGGCGAGATCCATATCGGCGGGGTGGGCCTGGCGCTCGGCTACGCCGGCGATCCG
CAGCGCACCGCCGAACGCTTCGTCCGTCACCCCGATGGCCGTCGCCTGTATCGCACCGGCGACCTCG
GCCGCTACCTGGCCGACGGCAGCATCGAGTTCCTCGGCCGCGAGGACGACCAGGTGAAGATTCGCGG
CCACCGCATCGAACTGGCCGAACTGGACGCCGCGCTGTGCGCTCATCCGCAGGTCAACCTGGCGGCC
ACCGTGGTGCTCGGCGAGACCCACGAGCGCAGCCTGGCCAGCTTCGTCACCCTGCATGCGCCGGTGG
AGGCTGGCGAGGATCCGCGTACGGCGCTCGACGCGGTGCGCCAGCGGGCGGCCCAGGCCTTGCGCCG
CGACTGGGGCAGCGAGGAGGGCATCGCCGCGGCGGTGGCCGCACTCGACCGTGCCTGCCTCGCCTCG
TTGGCCGCCTGGCTGGCCGGCAGCGGTCTGTTCGCCAGTGCGACGCCGCTGGACTTAGCCACCCTGT
GCCAGCGCCTGGGTATCGCCGAGGCGCGCCAGCGCCTGCTGCGCCACTGGTTGCGCCAACTGGAGGA
GGGCGGCTACCTGCGCGCCGAGGGCGAGGGCTGGCTGGGCTGCGCCGAGCGTCCCGCGCAGAGTCCG
GAGGACGCCTGGACGGCGTTCGCCGGCTGCGCGCCGGCGGCGCTCTGGCCGGCCGAGCTGGTCGCCT
ACCTGCGTGACAGCGCGCAATCCCTCGGCGAGCAACTGGCCGGGCGGATCAGCCCGGCGGCGCTGAT
GTTCCCGCAGGGCTCGGCGCGCATCGCCGAGGCCATGTACAGCCAGGGCCTGCATGCCCAGGCGCTG
CACGAGGCCATGGCCGAGGCCATCGCCGCCATCGTCGAGCGCCAGCCGCAACGGCGCTGGCGCCTGC
TGGAGCTTGGCGCCGGCACCGCCGCCGCCAGCCGCACGGTGATCGCCCGGTTGGCGCCGCTGGTGCA
GCGAGGGGCGGAGGTGGACTACCTGTTCACCGACGTTTCCAGCTACTTCCTCGCCGCCGCCCGCGAG
CGCTTCGCCGACCAGCCGTGGGTACGCTTCGGCCGCTTCGACATGAACGGCGATCTTCTCGACCAGG
GCGTGGCGCCGCACTCGGTGGATATCCTGCTCAGCTCCGGGGCCTTGAACAACGCGCTGGACACCCC
GGCGCTGCTCGCCGGCCTGCGCGAGTTGCTGAGCGCCGACGCCTGGCTGGTGATCCAGGAACTGACG
CGCGAGCACAACGAGATCAGCGTCAGCCAGAGCCTGATGATGGAAAACCCGCGCGACCTCCGCGACG
AGCGCCGCCAACTGTTCGTCCACACCGGGCAATGGCTGGAGTGGCTGGCGGCACAGGGTGGCGACCT
GGCTTGTGGGGTGGTGCCGCCGGGCAGCGCTCTCGACCTGCTTGGCTACGATGTCCTGCTGGCTCGC
TGCAAGACCGACCGCGCCCGCCTGGAGCCGGCCGAGCTGCTGGCCTTCGTCGAAGCGCGGGTGCCGC
GCTACATGCTCCCGGCGCAGTTGCGCGTGCTCGAACGCCTGCCGGTCACCGGCAACGGCAAGATCGA
CCGCAAGGCCCTGACCGGCTTTGCCCGCCAGCCCCAGGCGGACCTTCGGCATGGCGTCGCGCAGGCA
CCGGCCGACGAACTGGAGAATGCGCTGCTGGCACTCTGGCGGGAGGTGCTGGACAACCCGTCGCTGG
GCGTCGAGCAAGACTTCTTCGGGGCCGGCGGCGACTCGCTGTTGATCGCCCAGTTGATCGCCCGTTT
GCGCGAACGACTGGAAAGCGCCCGTCGGCATCCGTTCGATCGCCTGCTACGCTGGGCGCTCAGCCAG
CCGACGCCGCGCGGCCTGGCCGAACGCCTGCGCAGCGCGCCGGAAGAGGGCCGTGGGCCAGCCCTGG
CCGCGGCGCGCGGCGTCGCCCCGGCGCCGGCCGGCATGTCGCGCGCACCGCTCGCCGAGGGCGCGGT
GGCGCTCGACCCGCTGGTGCGCCTGGTGCCCGGCGAGGGCGTGCCGCGGGTGCTGGTCCACGAAGGC
CTCGGCACGCTACTGCCGTACCGCCCGCTGCTTCGCGCCCTGGGTGAGGGGCGGCCGTTGCTGGGGC
TGGCCGTGCATGACAGCGACGCCTACCTGGCGATCCCCGCCGAGCATCTCAACGCCTGCCTCGGCCG
CCGCTACGCCGAGGCGCTCCATCGCGCCGGGCTACGCGAGGTCGACCTGCTCGGCTACTGCTCCGGC
GGGCTGGTCGCCCTGGAGACCGCCAAGTCCCTGGTCCAGCGCGGGGTGCGCGTGCGCCAACTGGATA
TCGTCTCCAGCTACCGGATTCCCTACCGGGTGGACGACGAGCGCCTGCTGTTGTTCAGCTTCGCCGC
GACCCTCGGCCTGGATACCGCGGCGCTCGGCTTCCCCGCGCCGGAACGTCTCGGCCAGGCGGTGCAG
GCGGCGCTCGCGCAGACACCGGAGCGCCTGGTCGCCGAGGCGCTGGCGGGGCTGCCGGGCCTGGCCG
ATCTCGTCGCCCTGCGCGGCCGCGTGCTACAGGCGGCCAGCGGTAGCGCCGACGCCGTCAGCGTCGA
ACGCGACACCCTCTACCGGCTGTTCTGTCACTCGGTGCGTGCCAGCCAGGCCGAGGCGCCGGAGCCC
TACGTCGGCGCGCTGCGGCTGTTCGTGCCGGACGCCGGCAACCCATTGGTGCCGCGCTACGCCGAGG
CTCTGGAGACCCAATGGCGGGCCGCCGCGCTTGGCGCGTGCGGCATCCACGAGGTGCCCGGCGGGCA
CTTCGACTGCCTGGGCGAAGCCCTGGCGCAATCCTTGTCGAAACCCATGCCAGAGGAGGCGAGCCGA
TGA
[0065] The VIR16 protein (SEQ ID NO:32) encoded by SEQ ID NO:31 is
presented using the one-letter amino acid code in Table 18B.
TABLE-US-00034 TABLE 18B Encoded VIR16 protein sequence (SEQ ID NO:
32)
MSLGELLETCRSRRIELWSEAGRLRYRAPQGALDAGLAERLRAEREALLEHLEGGPGWRAEPDMA
HQRFPLTPVQAAYVLGRQAAFDYGGNACQLYAEYDWPADTDPARLEAAWNAMVERHPMLRAVIED
NAWQRVLPEVPWQRLTVHACAGLDEAAFQAHLERVRERLDHACAALDQWPVLRPELSIGRDACVL
HCSVDFTLVDYASLQLLLGEWRRRYLDPQWTAEPLEATFRDYVGVEQRRRQSPAWQRDRDWWLAR
LDALPGRPDLPLRVQPDTRSTRFRHFHARLDEAAWQALGARAGEHGLSAAGVALAAFAETIGRWS
QAPAFCLNLTVLNRPPLHPQLAQVLGDFTALSLLAVDSRHGDSFVERARRIGEQMFDDLDHPTFS
GVDLLRELARRRGRGADLMPVVFTSGIGSVQRLLGDGEAPRAPRYMISQTPQVWLDCQVTDQFGG
LEIGWDVRLGLFPEGQAEAMFDDFVGLLRRLAQSPRAWTDGDATEPVEAPPQALPGSARSIAAGF
AERALLTPDATAIHDAAGSYSYRQVAQHASALRRVLEAHGAGRGRRVAVMLPKSAAQLVAVIGIL
QAGAAYVPVDIRQPPLRRQAILASAEVVALVCLESDVPDVGCACVAIDRLAADSAWPPPPAAEVA
ADDLAYVIYTSGSTGTPKGVMLSHAAVSNTLLDINQRYGVDANDRVLGLAELSFDLSVYDFFGAT
AAGAQVVLPDPARGSDPSHWAELLERHAITLWNSVPAQGQMLIDYLESEPQRHLPGPRCVLWSGD
WIPVSLPTRWWRRWPDSALFSLGGATEAAIWSIEQPIRPQHTELASIPYGRALRGQSVEVLDARG
RRCPPGVRGEIHIGGVGLALGYAGDPQRTAERFVRHPDGRRLYRTGDLGRYLADGSIEFLGREDD
QVKIRGHRIELAELDAALCAHPQVNLAATVVLGETHERSLASFVTLHAPVEAGEDPRTALDAVRQ
RAAQALRRDWGSEEGIAAAVAALDRACLASLAAWLAGSGLFASATPLDLATLCQRLGIAEARQRL
LRHWLRQLEEGGYLRAEGEGWLGCAERPAQSPEDAWTAFAGCAPAALWPAELVAYLRDSAQSLGE
QLAGRISPAALMFPQGSARIAEAMYSQGLHAQALHEAMAEAIAAIVERQPQRRWRLLELGAGTAA
ASRTVIARLAPLVQRGAEVDYLFTDVSSYFLAAARERFADQPWVRFGRFDMNGDLLDQGVAPHSV
DILLSSGALNNALDTPALLAGLRELLSADAWLVIQELTREHNEISVSQSLMMENPRDLRDERRQL
FVHTGQWLEWLAAQGGDLACGVVPPGSALDLLGYDVLLARCKTDRARLEPAELLAFVEARVPRYM
LPAQLRVLERLPVTGNGKIDRKALTGFARQPQADLRHGVAQAPADELENALLALWREVLDNPSLG
VEQDFFGAGGDSLLIAQLIARLRERLESARRHPFDRLLRWALSQPTPRGLAERLRSAPEEGRGPA
LAAARGVAPAPAGMSRAPLAEGAVALDPLVRLVPGEGVPRVLVHEGLGTLLPYRPLLRALGEGRP
LLGLAVHDSDAYLAIPAEHLNACLGRRYAEALHRAGLREVDLLGYCSGGLVALETAKSLVQRGVR
VRQLDIVSSYRIPYRVDDERLLLFSFAATLGLDTAALGFPAPERLGQAVQAALAQTPERLVAEAL
AGLPGLADLVALRGRVLQAASGSADAVSVERDTLYRLFCHSVRASQAEAPEPYVGALRLFVPDAG
NPLVPRYAEALETQWRAAALGACGIHEVPGGHFDCLGEALAQSLSKPMPEEASR
MUT17
[0066] A Pseudomonas bacterial mutant (MUT17) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding, putative ATP-binding component
of the ABC transporter, pchH (PA4223). This gene encodes the VIR17
nucleic acid (SEQ ID NO:33) shown in Table 19A. TABLE-US-00035
TABLE 19A VIR17 Nucleotide Sequence (SEQ ID NO: 33)
GTGACCCCGGTGCTGTGGCGCCTGCTGCGCACCTATCGCTGGCGGCTGGCGGCGGCCATGGGGTTGC
AGGCCCTGGCCGGGCTCTGCTCGCTGTTGCCCTGGATGCTTCTCGCCTGGCTCGCCGAGCCGCTGGC
GCGCGGCCAGGCGCAGCCGGCCCTGCTGGCCCTGGTGCTGCTGGCGGTGCTGGCCTGGCTGGGCTGC
CAGGCGCTGCCCGCGCACCTGGCCCACCGGGTCGACGCGGACCTCTGCAACGACCTGCGCCTGCGCC
TGCTGGCGCACCTGCAACGGCTGCCGCTGGACTGGTTCGGTCGCCAGGGCCCGGACGGCGTGGCGCG
CCTCGTGGAGCAGGACGTGCGGGCCCTGCACCAACTGATCGCGCACGCTCCCAACGATCTCAGCAAC
CTGTTGGTGGTGCCGCTCGTCGCGTTGCTCTGGCTGGCCTGGCTGCACCCCTGGCTGCTGCTGTTCT
GCCTGCTGCCGCTGGTGCTGGCCGCCGCCGGCTTCCTGCTGCTGCGCTCGGCGCGCTACCGCGACCT
GGTGCTGCGGCGCAACGCCGCGCTGGAAAGGCTCTCGGCGGACTATGGCGAATTCGCCCACAACCTG
CTGCTGGCCCGACAGTACCCCGGCGCCGGCATACAACAGGGCGCCGAGGCGTCGGCGGCGGCCTTCG
GCGAAGCGTTCGGCGCCTGGGTGAAGCGGGTCGGCCACCTCGCCGCGCTGGTCTACGTGCAGTTGTC
GACGCCCTGGCTGCTGGCCTGGGTCCTGCTCGGCGCGCTGGCCCTGGATGCCCTCGGCGTGCCGCTG
GCGCTCGGCCAGGCCTGTGCCTTCCTGCTCCTGCTGCGGGCCTTGGCTGCCCCGGTGCAGGCGCTCG
GCCACGCCGGCGACGCGCTGCTGGGCGCGCGCGCCGCCGCCGAGCGCCTGCAGCAGGTGTTCGACCA
GGCGCCGCTGGCCGAGGGCCGCTCGACCCGCGAGCCGGTCGATGGCGCGGTGGCGCTGCACGGCCTG
GGCCATGCCTATGAAGGCGTGGAGGTCCTGGCCGATATCGATCTGGAGCTGGAGGATGGCAGCCTGG
TGGCCCTGGTCGGTCCCTCGGGCTCCGGCAAGAGCACCCTGCTGCACCTGCTGGCGCGCTACATGGA
CGCGCAGCGCGGCGAACTGGAGGTTGGCGGCCTGGCACTGAAGGACATGCCTGATGCCGTGCGCCAT
CGGCATATCGCGCTGGTCGGCCAGCAGGCGGCGGCGCTGGAGATATCCCTGGCCGACAACATTGCCC
TGTTCCGCCCCGATGCCGATCTCCAGGAGATTCGCCAGGCGGCCCGTGACGCCTGCCTCGACGAGCG
CATCATGGCCCTGCCGCGTGGCTACGACAGCGTGCCGGGACGCGACCTGCAACTGTCCGGCGGCGAA
CTGCAACGACTGGCCCTGGCCCGTGCGCTGCTATCGCCGGCGAGCCTGTTGCTGCTCGACGAGCCAA
CCTCGGCGCTGGATCCGCAGACCGCCCGGCAGGTCCTGCGCAACCTGCGCGAACGCGGCGGTGGCCG
GACCCGGGTGATCGTCGCCATCGTCTGGCCGAAGTCAGCCGATGCCGACCTGATCCTGGTGCTGGTC
GCTGGCCGTCTGGTCGAACGCGGCGAGCACGCGGCGCTGTTGGCGGCGGACGGCGCCTATGCGCGCT
TGTGGCGTGAACAGAACGGCGCGGAGGTGGCGGCATGA
[0067] The VIR17 protein (SEQ ID NO:34) encoded by SEQ ID NO:33 is
presented using the one-letter amino acid code in Table 19B.
TABLE-US-00036 TABLE 19B Encoded VIR10 protein sequence (SEQ ID NO:
34)
MTPVLWRLLRTYRWRLAAAMGLQALAGLCSLLPWMLLAWLAEPLARGQAQPALLALVLLAVLAWL
GCQALAAHLAHRVDADLCNDLRLRLLAHLQRLPLDWFGRQGPDGVARLVEQDVRALHQLIAHAPN
DLSNLLVVPLVALLWLAWLHPWLLLFCLLPLVLAAAGFLLLRSARYRDLVLRRNAALERLSADYG
EFAHNLLLARQYPGAGIQQGAEASAAAFGEAFGAWVKRVGHLAALVYVQLSTPWLLAWVLLGALA
LDALGVPLALGQACAFLLLLRALAAPVQALGHGGDALLGARAAAERLQQVFDQAPLAEGRSTREP
VDGAVALHGLGHAYEGVEVLADIDLELEDGSLVALVGPSGSGKSTLLHLLARYMDAQRGELEVGG
LALKDMPDAVRHRHIALVGQQAAALEISLADNIALFRPDADLQEIRQAARDACLDERIMALPRGY
DSVPGRDLQLSGGELQRLALARALLSPASLLLLDEPTSALDPQTARQVLRNLRERGGGRTRVIVA
HRLAEVSDADLILVLVAGRLVERGEHAALLAADGAYARLWREQNGAEVAA
[0068] The role of VIR17 in virulence was confirmed using phage to
retransduce this mutation into the wild-type PT894 strain where
attenuated virulence was again observed in the Dictyostelium growth
assay compared to an isogenic bacterial strain.
MUT18
[0069] A Pseudomonas bacterial mutant (MUT18) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding the putative ATP-binding
component of ABC transporter, pchI (PA4222). This gene encodes the
VIR18 nucleic acid (SEQ ID NO:35) shown in Table 20A.
TABLE-US-00037 TABLE 20A VIR18 Nucleotide Sequence (SEQ ID NO: 35)
ATGACCCTGTTCGAACGAATGCGTGCGCTGCCCGAAGACTGCCGTGCCGCGTTGCGCCGGGCGAGCG
CCTGGGCGGTCCTGGCGGCGCTGCTGGACGCCGCTTGCGGCGTATTGCTGGTGCCGTTGGTCGAGGC
CTGGTTCGCCGAAGGCGCGTTGCCCTGGCGCTGGGTCGCCGCGTTGCTCGGCTTGAGCCTGGCGCAG
GCGCTGTTGCAGTACCTGGCCCTGCGTCGCGGTTTCGCCGCCGGCGGCTCGCTGGCGGCTGGACTGG
TGCGCAGCCTGGTGGCGCGCTTGCCGCGCCTGGCGCCGCCGGCGCTGCGCCGGGTCGCGCCGGCCGA
AGGCCTGCTGCGCGGCCCGGTGATGCAGGCGATGGGCATTCCGGCGCACCTGCTGGGGCCGCTGATC
GCCGCGTTGGTGACGCCGCTCGGGGTGATCCTCGGGCTGTTCCTGATCGACCCGTCCATCGCCCTCG
GCCTGCTCCTTGCTGGTGCCTTCCTCGCCGCGCTGTTGCGCTGGAGCGGGCGGCGCAATCTGGCGGC
GGAGGATGCCCGGCTGGCCGCCGAGCGCGACGCCGCACGGCAGTTGCAGGCGTTCGCCGAACGCCAG
CCACTGCTGCGCGCGGCGCAGCGCGAAAGCGTCGCCCGCCAGGGGCTGGAAGAGGCCTTGCGCAGTC
TCCACCGCAGCACCCTGGATCTGTTGCGGCGCAGCCTGCCCAGCGGCCTCGGCTTCGCCCTGGCGGT
GCAGGCGGCGTTCGCCTTCGCCCTGCTCGGCGGCGCCTGGGCGGTGGAGCGGCAATGGCTGGACCGC
GCTCGGCTGGTGGCCGTGCTGGTGCTGCTGGTGCGCTTCATCGAGCCGCTGGCCCAGCTCACCCATC
TCGACCAGGCGTTGCGCGGCGCCTGGCAGGCGCTGGATACCCTGCTGCGGGTTTTCGCCCTGGCTCC
GCTGCGCAGCCCCGAGCCGGGCGAGCGGCCGCACGACGCCAGCCTGGCGGCCGAGGCCGTGGAATTG
CGCCTGGAAGATGGCCGCGCCTTGCTCGAGGACATTTCCCTGAGGCTGGAGCCGGGTTCGCTGAACG
TCCTCGTCGGACCCTCCGGGGCCGGCAAGAGCAGCCTGCTGGCGCTGCTCGGGCGGCTCTACGACGT
CGATGCCGGGCGTGTCCTGCTGGGTGGCGTGGATATCCGCCGGTTGAGCGAAACGACCCTCGCCGCC
AGTCGTAACCTGGTGTTCCAGGACAACGGCCTGTTCCGCGGCAGCGTTGCCTGGAACCTGCGCATGG
CGCGAGCGGACGCCGATCTCGAAGCGCTGCGCGAGGCGGCGCGGGCGGTTGGCCTGCTGGAAGAGAT
CGAGGCCTGGCCGCAGGGCTGGGACAGCGACGTCGGTCCCGGCGGCGCGCTGCTGTCCGGCGGCCAG
CGGCAACGCCTGTGCCTGGCTCGCGGGCTGCTCTCGACGGCGCCGTTGCTGCTGCTCGACGAGCCCA
CCGCCAGCCTCGACGCCGCCAGCGAGGCGCAGGTGCTGCGCAGCCTGCTCGGGTTGCGCGGCCGGCG
CACCCTGCTGGTAGTGACCCACCGCCCGGCGCTGGCGCGTCAGGCCGACCAGGTACTGCTGCTGGAG
GAGGGGCGCCTGCGCCTCACCGGACTTCACGCCGATCTGCTCGTCCGGGACGACTGGTATGCCGGTT
TCGTCGGGCTGGCGGGCGAGGAAAGTTCCGCGACGGTCGTGGATCGATAG
[0070] The VIR18 protein (SEQ ID NO:36) encoded by SEQ ID NO:37 is
presented using the one-letter amino acid code in Table 20B.
TABLE-US-00038 TABLE 20B Encoded VIR18 protein sequence (SEQ ID NO:
36)
MTLFERMRALPEDCRAALRRASAWAVLAALLDAACGVLLVPLVEAWFAEGALPWRWVAALLGLSL
AQALLQYLALRRGFAAGGSLAAGLVRSLVARLPRLAPPALRRVAPAEGLLRGPVMQAMGIPAHLL
GPLIAALVTPLGVILGLFLIDPSIALGLLLAGAFLAALLRWSGRRNLAAEDARLAAERDAARQLQ
AFAERQPLLRAAQRESVARQGLEEALRSLHRSTLDLLRRSLPSGLGFALAVQAAFAFALLGGAWA
VERQWLDGARLVAVLVLLVRFIEPLAQLTHLDQALRGAWQALDTLLRVFALAPLRSPEPGERPHD
ASLAAEAVELRLEDGRALLEDISLRLEPGSLNVLVGPSGAGKSSLLALLGRLYDVDAGRVLLGGV
DIRRLSETTLAASRNLVFQDNGLFRGSVAWNLRMARADADLEALREAARAVGLLEEIEAWPQGWD
SDVGPGGALLSGGQRQRLCLARGLLSTAPLLLLDEPTASLDAASEAQVLRSLLGLRGRRTLLVVT
HRPALARQADQVLLLEEGRLRLSGLHADLLVRDDWYAGFVGLAGEESSATVVDR
[0071] The role of VIR18 in virulence was confirmed using phage to
retransduce this mutation into the wild-type PT894 strain where
attenuated virulence was again observed in the Dictyostelium growth
assay compared to an isogenic bacterial strain.
MUT19
[0072] A Pseudomonas bacterial mutant (MUT19) was made by
transposon insertion in a P. aeruginosa wild-type strain PT894. In
the Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as a gene cluster encoding the P. aeruginosa
serotype 09 putative O-antigen biosynthesis pathway (VIR19). The
insertion site nucleic acid sequence identifying the VIR19 gene in
MUT19 is shown in Table 21. TABLE-US-00039 TABLE 21 MUT19
Transposon insertion Site (SEQ ID NO: 37)
CTCTTTCAGCCGCACGCGGCGCACCTCGTGTGTGATCAGTGAGTGGTTTGCAACTGCGGGTCAAG
GATCTGGATTTCCCTCACANGTNCGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCT
TGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGGGNNAATTGATCCGGTGGATGACC
TTTTGAATGACCTTTAATAGATTATATTACTAATTAATTGGGGACCCTANAGGTCCCCTTTTTTA
TTTTAAAAATTTTTTCACAAAACGGTTTATTTNCATAAAGCTTGCTCAATCAATCACCNTATCCN
CGGGAATTCGGCCTAGGCGGCCAGATCTGATCAAGAGACAGACCTCCAGCTTTGCATCCGGAGCG
ACCACACGAGCGAGGTCAGTCACTTTCATCGAAGGAATTTTCTTGACATAGATCTCACCACCTTC
CATGTCCTCAAAGGCATGCCACACTAACTCGACGCCCTCCTCCAAAGAAATCATGAACCGGGTCA
TCCGCTCATCAGTGATAGGCAAGACGCCCTTGTCCTTG
[0073] The role of this cluster in virulence was confirmed using
phage to retransduce this mutation into the wild-type PT894 strain
where attenuated virulence was again observed in the Dictyostelium
growth assay compared to an isogenic bacterial strain.
[0074] B. Attenuated Klebsiella Mutants
MUT20
[0075] A Klebsiella bacterial mutant (MUT20) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding a hypothetical transcriptional
regulator in met G-dld intergenic region (VIR20). The insertion
site nucleic acid sequence identifying the VIR20 gene in MUT20 is
shown in Table 22. TABLE-US-00040 TABLE 22 MUT20 Transposon
Insertion Site (SEQ ID NO: 38)
ACGCAGGATATCTTCTTCATCAAATTGTCGATGCCCGCCTTCGCTACGCTGCGGTTTCAGTAGACCG
TAACGACGCTGCCAGGCGCGCAGTGTGACCGGATTGATTCCGCAACGTTCGGCGACTTCACCGATAC
TGTAAAACGCCATAGCAGCCTCACATCAACCTGATACCTTAATACCTAAACTAACGAATTCAGGCAT
CCTGTACAACTCTATTTTCTTGTACAGATAAAGATATCAGGTTGCGGCTCACAGCGCCCGGGAAAAA
AGATGAAAAAATGTTTAGCTGATTTCGCGGTGGTTCATTTTTTCTCCGGCCATGCGACGGCGGGTAG
GCCCCCCAGGCGCGCGCTGGCGAACAAATTGCCCTGAAACTGTGAAATACCGGCTGATTCCAGCCAC
ATCCACTCTTCAGCACGCTCAACGCCGACGGCTGAGACCGCAATCTCCAGAGAAGTACAGCATTTGA
TAATCGCCTG
MUT21
[0076] A Klebsiella bacterial mutant (MUT21) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding .cndot.-cystathionase (VIR21).
The insertion site nucleic acid sequence identifying the VIR21 gene
in MUT21 is shown in Table 23. TABLE-US-00041 TABLE 23 MUT21
Transposon Insertion Site (SEQ ID NO: 39)
GACCATGTGCTGATGACCAATACCGCCTATGAGCCAAGCCAGGACTTTTGTACCAAAATTCTCGCCA
AACTCGGCGTCACCACCAGCTGGTTCGATCCCTTAATCGGCGCCGATATCGCCCGTCTGGTTCGCCC
TGAGACCCGCGTGGTGTTCCTCGAATCGCCCGGCTCGATCACCATGGAAGTGCACGATGTGCCGGCG
ATAGTCGCCGCCGTGCGTCAGGTCGCCCCGGAAGCGATTATCATGATCGATAACACCTGGGCGGCGG
GGATCCTGTTTAAAGCCCTGGATTTTGGCATTGATATTTCCATTCAGGCAGGCACCAAATACCTGAT
CGGCCATTCCGACGCCATGGTGGGCACCGCGGTGGCGAACGCGCGCTGCTGGCCGCAGCTGCGTGAA
AATGCCTACCTGATGGGGCAAATGCTGGACGCCGATACTGCCTATATGACCAGCCGCGGCCTGCGAA
CCCTGGGCGTGCGCCTGCGTCAGCATCATGAAAGCAGCCTGCGCATC
MUT22
[0077] A Klebsiella bacterial mutant (MUT22) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as ribosome binding factor A (VIR22). The insertion
site nucleic acid sequence identifying the VIR22 gene in MUT22 is
shown in Table 24. TABLE-US-00042 TABLE 24 MUT22 Transposon
Insertion Site (SEQ ID NO: 40)
CTTTTGGCCCCTTTTTTGTCTTTATTCTGGAGAACTTATTATGGCGAAAGAATTTGGTCGCCCGCAG
CGTGTGGCCCAGGAGATGCAAAAAGAGATTGCCATCATCCTGCAGCGTGAAATTAAAGATCCGCGTC
TGGGCATGATGACCACCGTTTCCGGTGTGGAAATGTCCCGTGACCTGGCCTATGCCAAGGTGTATGT
CACCTTCCTTAACGACAAAGATGAAGCCGCGGTGAAAGCGGGCATCAAAGCGCTGCAGGAAGCTTCT
GGCTTTATCCGCTCTCTGCTGGGGAAAGCGATGCGTCTGCGCATCGTACCGGAACTGACTTTCTTCT
ACGACAACTCACTGGTGGAAGGGATGCGTATGTCCAACCTGG
MUT23
[0078] A Klebsiella bacterial mutant (MUT23) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding aspartokinase/homoserine
dehydrogenase (VIR23). The insertion site nucleic acid sequence
identifying the VIR23 gene in MUT23 is shown in Table 25.
TABLE-US-00043 TABLE 25 MUT23 Transposon Insertion Site (SEQ ID NO:
41)
GCCCAGCCCGCTTTCCCGCTTGCCCAGTTAAAAGCCTTCGTGGAGCAGGAATTTGCTCAGATTAAGC
ATGTTCTGCACGGCATCAGCCTGCTGGGTCAGTGCCCGGACAGCGTCAATGCCGCGCTGATCTGCCG
CGGCGAAAAGCTCTCCATCGCCATCATGGCGGGTCTGCTGGAAGCCCGTGGACACAAAGTCAGTGTC
ATTAACCCGGTCGAAAAACTGCTCGCCGTGGGTCACTATCTGGAATCCACCGTCGATATCGCCGAAT
CCACCCGCCGCATTGCCGCCAGCCAGATCCCGGCAGACCATATGATCCTGATGGCCGGGTTTACCGC
CGGCAATGAGAAAGGCGAGCTGGTGGTGCTGGGGCGTAACGGCTCCGACTACTCGGCTGCGGTACTG
GCCGCCTGCCTGCGCGCTGACTGCTGCGAAATCTGGACCGATGTCGACGGAGTGTACACCTGCGATC
CGCGTCAGGTGCCGGATGCGCGCCTGCTGAAATCGATGTCTTATCAGGAGGCGATGGAGCTCTCCTA
CTTTGGCGCGAAAGTGCTGCACCCGCGCACCATTGCCCCTATCGCCCAGTTCCAAATCCCATGCCTG
ATTAAAAATACCGGCAACCCCC
MUT24
[0079] A Klebsiella bacterial mutant (MUT24) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding cystathione .cndot.-synthetase
(VIR24). The insertion site nucleic acid sequence identifying the
VIR24 gene in MUT24 is shown in Table 26. TABLE-US-00044 TABLE 26
MUT24 Transposon Insertion Site (SEQ ID NO: 42)
GGCGCAGCGTCTGCTCGTCACCGTCAAGCTCGAAGCTTAACATTGCGCCAAAACCTTTTTGCTGACG
CGCCGCAATTTCATGCCCCTGGTTTTCCGGCAGCGATGGATGATACAGCTTTTTCACCAGCGGCTGG
GTTTTCAGATACTCAACGATCGCCAGGGCATTTCGCTGCGCCACTTCCATCCGTGGAGACAGCGTCC
GCAGCCCGCGCAACAGCAGATAGCTGTCGAAGGCGCTGCCGGTGACGCCAATATTATTCGCCCACCA
TGCCAGTTCGGTGACAGTTGCCGGATCTTTGGCAATCACCACCCCGGCCACCACATCGGAGTGACCA
TTGAGGTATTTGGTACAGGA
MUT25
[0080] A Klebsiella bacterial mutant (MUT25) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding
phosphoribosylformylglycinamidine synthase (VIR25). The insertion
site nucleic acid sequence identifying the VIR25 gene in MUT25 is
shown in Table 27. TABLE-US-00045 TABLE 27 MUT25 Transposon
Insertion Site (SEQ ID NO: 43)
GTTGCGTCCCAGGCGCGTAAACGCATCCTGCAGGTAGTCAATTTCGTCGTCGGCCAGCGCCAGACCC
AGACGGAGGTTGGCGTCAATCAGCGCCTGACGCCCTTCGCCCAGCAGGTCGACGCTGGTGACCGGCG
TCGGCTGATGGTGAGCGAACAGCTTCTCGCCCGCTTCCAGCTCGTCGAAGACGCTCTCCATCATGCG
GTCATGCAGCTCCGCCGCCACCGCGGCCCACTGCGCTTCGGTCAGGGTTGAGGCTTCAACGTAATAC
GCCACGCCGCGCTCAAGACGCACAACCTGCGCCAGACCGCAGTTGTGAGCGATATCGGTAGCTTTAG
AAGACCAGGGAGAGATGGTGCCAGGGCGAGGGGTCACGAGCAGTAATTTACCGGTCGGGGTATGGCT
GCTTAAGCTCGGGCCATACTGAAGCAGTCGCGCCAGGCGCTCGCGATCGTCAGCGCTCAGCGGGCCG
TTCAGATCGGCAAAATGAATATATTCGGCAT
MUT26
[0081] A Klebsiella bacterial mutant (MUT26) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding homoserine transsuccinylase
(VIR26). The insertion site nucleic acid sequence identifying the
VIR26 gene in MUT26 is shown in Table 28. TABLE-US-00046 TABLE 28
MUT26 Transposon Insertion Site (SEQ ID NO: 44)
GTATTGGCATCGTACTCCTGGGCTGGCCGGTGACAAAGGCGATGCGCTTATCTTTGCTGGCGAACAA
ATACGCATCGCCCTCTTCCGTCTCCGCGAGGATCTCGAGATCGGTATAGTCGCGAATAAGTCCGGCC
GGAAAATCAGCATAGCGTGAGTGCGGGGCCAGGAAAGAGTCGTCGAAACCGCGGGTCAGTAAGGCGT
GCGGATGAAGAATATGGTGTTCATAGACGCCGGAAATCTTTTCGGCGCGGGTCTGCTTGGGAATGCC
GTACAGAATGTTCAGCGCGGCCTGAACCGCCCAACAGACGAACAGCGTCGAAGTGACGTGATCCTTG
GCCCACTCCAGCACCTGTTTGATCTGCGGCCAGTAAGCAACATCGTTAAACTCAACCAGGCCTAAAG
GAGCGCCGGTAACAATCAGGCCGTCAAAGTTCTGATC
MUT27
[0082] A Klebsiella bacterial mutant (MUT27) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding 3'-phosphoadenosine
5'-phosphosulfate reductase (VIR27). The insertion site nucleic
acid sequence identifying the VIR27 gene in MUT27 is shown in Table
29. TABLE-US-00047 TABLE 29 MUT27 Transposon Insertion Site (SEQ ID
NO: 45)
GAGGTTCATATGTCCGTACTCGATCTAAACGCGCTTAATGCATTGCCGAAAGTGGAACGCATTCTGG
CACTCGCGGAAACCAACGCCCAACTGGAAAAGCTTGACGCCGAAGGGCGTGTGGCGTGGGCGCTGGA
AAATCTGCCGGGAAACTATGTGCTGTCGTCGAGCTTTGGCATTCAGGCGGCGGTAAGTTTGCATCTG
GTGAATCAGATCCGCCCGGACATTCCGGTGATCCTCACCGATACCGGCTACCTGTTCCCGGAAACCT
ATCAGTTTATTGACGAGCTGACGGACAAG
MUT28
[0083] A Klebsiella bacterial mutant (MUT28) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding Sfi protein (VIR28). The
insertion site nucleic acid sequence identifying the VIR28 gene in
MUT28 is shown in Table 30. TABLE-US-00048 TABLE 30 MUT28
Transposon Insertion Site (SEQ ID NO: 46)
TGTTAAAGCGTGCGTTCTACAGCCTGTTAGTCCTGCTCGGCCTGCTGCTGTTGACCGTGCTGGGCCT
TGACCGCTGGATGAGCTGGAAAACCGCGCCCTATATCTATGATGAACTGCAGGACCTGCCCTACCGT
CAGGTCGGTGTGGTGCTGGGCACCGCCAAATATTACCGCACCGGCGTCATCAATCAGTATTACCGTT
ACCGCATCCAGGGTGCGCTGAACGCCTACAACAGCGGCAAGGTCAACTATCTCCTGCTGAGCGGCGA
TAATGCTCTGCAAAGCTACAATGAACCGATGACCATGCGTCGGGACCTGATTAAAGGCGGCGTCGAT
CCCGCGGATATCGTACTGGACTATGCCGGTTTCCGTACCCTCGACTCGATCGTCCGTACCCGGAAAG
TGTTCGACACCAACGACTTCATTATCATCACCCAGCGCTTCCACTGCGAACGGGCGCTGTTTATCGC
CCTGCATATGGGGATCCAGGCCCAGTGCTACGC
MUT29
[0084] A Klebsiella bacterial mutant (MUT29) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding transcriptional activator
protein LysR (VIR29). The insertion site nucleic acid sequence
identifying the VIR29 gene in MUT29 is shown in Table 31.
TABLE-US-00049 TABLE 31 MUT29 Transposon Insertion Site (SEQ ID NO:
47)
CGCTGAACCTCCTCAAACAAACGCAGGCCCTGCACCTGTCGGCTGCAGGCGACCAGCGTGGATCCGC
TCAAACAGCTGCAGGCCGAGCACCTTCTCAAAGCGCGCCAGCTCGCGGCTGACCGTGGGTTGCGAGG
TGTGCAGCATCCGCGCCGCTTCGGTCAGGTTGCCGGTGGTCATCACCGCGTGAAAGATTTCGATATG
ACGCAAATTGACGGCTGGCATGCGGTCTCCGTGAGGCTCGGCTGGAACCATATCATTTTTGCATAGA
GTCGCGATAAAACGATATTTTTTATTCGTCTGTCACTGTGGCGTAATCAGAAAAAACAGCGACCAAC
ACACGCACTGCACCGGAGTTCTTATGCCACACTCGCTTTACGCCACCGATACTGACCTGACCGCGGA
CAACCTGCTGCGCCTGCCGGCGGAATTTGGCTGCCCGGTCTGGGTCTATGATGCGCAGATTATTCGC
CGCCAGATAGCCCAGCTCAGCCAGTTTCGAC
MUT30
[0085] A Klebsiella bacterial mutant (MUT30) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding TrpD (VIR30). The insertion
site nucleic acid sequence identifying the VIR30 gene in MUT30 is
shown in Table 32. TABLE-US-00050 TABLE 32 MUT30 Transposon
Insertion Site (SEQ ID NO: 48)
GGCTTCCACCCAAATCGCTTTGTCGGCAACGATTTTTGCTAAAACGGCTTTGCATTCTTTACCCTCT
TGCCCGCTAAGTGTCGGTCACTCTGTCATAGGCCGCGCGCTGCTGCAGCACATCCAGTACCTGCTGA
GCGTTAGCTTTCAGATCTTCATGCCCGTGTAAACGCATCAATATGGCGACGTTGGCGGCGACGGCGG
CTTCGTGAGCGGCTTCACCTTTACCTTG
MUT31
[0086] A Klebsiella bacterial mutant (MUT31) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding N-acetylglucosamine-6-phosphate
deacetylase (VIR31). The insertion site nucleic acid sequence
identifying the VIR31 gene in MUT31 is shown in Table 33.
TABLE-US-00051 TABLE 33 MUT31 Transposon Insertion Site (SEQ ID NO:
49)
TGGCTCAACGCTGCTCAGTGGTGCGAGGTGTCACTTTGGTGATCACATCGGCGTTGTCTGCACAGTG
AAATCAGATCCAGCGCCGCGTCCGGTTTTACGCACGTAGTCCGGATTGTGGGTGCCTTTCTTAACGA
TATTCAGCCACGGCCCTTCGAGATGCAGGCCCAGCGCCTGGTTCGGATGTTTTTGCAGATATTCGCG
CATCACGCGCACGCCTTGCTTCATCAGATCGTCGCTGGAGGTAATCAGCGTCGGCAGGAAGCTGGTG
CAGCCTGAGCGTTCGTTGGCCTTCTGCATGATCTCCAGCGTTTCGACAGTGACCGCCTCTGGGCTGT
CGTTAAACTGCACGCCGCCGCAGCCGTTGAGCTGGACGTCGATAAAACCGGGGGCGATTATTGCGCC
GTTGACTGAGCGCTGCTCGATGTCAGACGGCAAATCTGCCAGCGGACAAAGACGTTCGATAAAG
MUT32
[0087] A Klebsiella bacterial mutant (MUT32) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding WaaQ (VIR32; Regue et al. J.
Bacteriol. 183(12): 3564-73, 2001). The insertion site nucleic acid
sequence identifying the VIR32 gene in MUT32 is shown in Table 34.
TABLE-US-00052 TABLE 34 MUT32 Transposon Insertion Site (SEQ ID NO:
50)
TTAAGCACCATATCGTACCGCTGCTGGCGCAGCGTCTGAATGAGCTGCCATTGCATCTTCAGCTGAT
ACCTTTTTCCCTGGCTTTTTCCAGCGGCGATCGAGACCATAAATATGGTGGATATCGGGGTTGGCTG
CGAGCATATCCCGGGTCTCTTCATACAACAGGACATCCACGCTGGCGGCGGGGTACTGCTGTTTCAG
CGCGTGAATAAGCGGCGTGATCAGCAGCATGTCGCCATGATGGCGCAGCTTAATGACCAGGATCCGC
GCCGGGTTCAACGGGCCGCGGGAGAGGGTTTCAGGCGTCATACTCTGTTCTTCATCCAGGATAAGGG
TTCCGATTCTAGGGGATCAGACAGATTGAGAGAAGCGTTGTATTGCTCTACCATGACCCGATACGTA
TGGCCTGAGGACGTTTTCGTGCACAATCCCGCAATTTTCTCATCACGAT
MUT33
[0088] A Klebsiella bacterial mutant (MUT33) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding 2-isopropylmalate synthase
(VIR33). The insertion site nucleic acid sequence identifying the
VIR33 gene in MUT33 is shown in Table 35. TABLE-US-00053 TABLE 35
MUT33 Transposon Insertion Site (SEQ ID NO: 51)
CACTCAGGCTTGCCTGTAACGCTTGTTCGCCATCACGTAAGGTCGTATCGAAAATAATGACTTGCTG
GCTCATGGTTTGGATCCTTAGTCTGTGTCCTGGCGCCTTGTTGACGAGCATAAAAAAACCCGCGCCA
AGGCGCGGGTTTTATAGTCTTGCTGGAAGATGACTTAACGCTGAACGTCGCCCAACAGCCTACCGAG
CAAATGGCATGCGTTTAGTAGTAGTAGGCTGGTGATACGAGCGGTGCGAATCATTGCGTCAAACTCC
AGATGAAATCGTTATGCTTTTAGAGTTACTGGATAGCCGTTTTAAAGTCAACCCCTGGCATGGAAAA
AGCGTTTTGGGCTGACTAAATGAATTAGCAAAATGTGCTGATGTAAGCCCCATTTTGCCGAAGATCC
TATTTTGGACCGAAGGCGGTTTATCCCCAATTTGTTTCATTTGAAAAA
MUT34
[0089] A Klebsiella bacterial mutant (MUT34) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding histidinol dehydrogenase
(VIR34). The insertion site nucleic acid sequence identifying the
VIR34 gene in MUT34 is shown in Table 36. TABLE-US-00054 TABLE 36
MUT34 Transposon Insertion Site (SEQ ID NO: 52)
CGCTGAACCGCTATCCGGAGCCGCAGCCGAAGTGCCGTGATTGAGAGCTACGCCCGCTACGCCGAGG
TCAAACCGGAGCAGGTGCTGGTCAGCCGCGGCGCCGACGAAGGCATCGAGCTGCTGATCCGCGCCTT
CTGTGAGCCCGGCGAAGACGCGGTGCTCTACTGCCCGCCGACCTACGGCATGTACAGCGTCAGCGCC
GAGACCATCGGCGTCGAGTGCCGCACCGTGCCGACGCTGGCCAGCTGGCAGCTCGACCTGCCGGGCA
TCGAAGCGCGGCTGGACGGCGTGAAGGTGGTGTTTGTCTGCAGCCCGAACAACCCGACCGGGCAGAT
TATCGACCCGCAGTCGATGCGCGACCTGCTGGAGATGACCCGCGGCAAAGCCATCGTGGTGGCCGAC
GAAGCCTATATTGAATTCTGCCCGCAGGCGACGCTCGCCGGCTGGCTCAGCGACTATCCGCACCTGG
TGGTGCTGCGCACGCTGTCCAAAGCCTTCGCCCTCGCCGGCCTGCGCTGCGGCTTCACCCTCGCCAA
CGCCGAGGTGATTAACGTGCTGCTGAAAGTGATCGCCCC
MUT35
[0090] A Klebsiella bacterial mutant (MUT35) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding UDP-galactopyranose mutase
(VIR35; Clarke et al., J. Bacteriol., 177: 5411-18, 1995). The
insertion site nucleic acid sequence identifying the VIR35 gene in
MUT35 is shown in Table 37. TABLE-US-00055 TABLE 37 MUT35
Transposon Insertion Site (SEQ ID NO: 53)
CGTATATTTCATCGTACAGAAACCGTAAACACAGGCATTGGCTGATTTTCAGTGAGTGAATTTAAAT
AGACTTCTGCCGTTTTCAATGCTTCGGCGATGGTCACATCCATATCAAGGTAACGGTAGGTTCCAAG
ACGACCGACAAAAGTGATGTTGGTTTCATTCTCGGCCAATGACAAATATTTTTCAAGAAGAGCCATT
TCTCCCATCTGGCGAATAGGATAGTAAGGAATATCATTTTCTTCACAAGCACGGCTATACTCTTTAT
AACAAACAGAGCCGTCGTGTTGTTCCCAGGGAGAAAAATATTTATGTTCAGTGATGCGAGTATAGGG
CACATCCACAGAACAGTAGTTCATCACTGCGCATCCCTGG
MUT36
[0091] A Klebsiella bacterial mutant (MUT36) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the scene encoding O-antigen export system
permease protein rfba (VIR36; Bronner et al., Mol. Microbiol., 14:
505-19, 1994). The insertion site nucleic acid sequence identifying
the VIR36 gene in MUT36 is shown in Table 38. TABLE-US-00056 TABLE
38 MUT36 Transposon Insertion Site (SEQ ID NO: 54)
GTACGCCGATTTTATATGCGTCTGATATGATTCCGGAAAAATTTAGCTGGATAATTACCTACAATCC
GCTAGCGAGTATGATTCTTAGTTGGCGTGATTTATTCATGAATGGGACTCTTAATTTTGAGTATATT
TCTATACTCTATTTTACGGGAATTATTTTGACGGTTGTCGGTTTGTCTATTTTCAATAAATTAAAAT
ATCGATTTGCAGAGATCTAAAAGTGCGCTATAAGAGCAGCATGCTAGGCTATTTATGGTCAGTAGCA
AATCCATTGCTTTTTGCCATGATTTACTATTTTATATTTAAGCTGGTAATGAGAGTACAAATTCCAA
ATTATACAGTTTTCCTCATTACCGGCTTGTTTCCGTGGCAATGGTTTGCCAGTTCGGCCACTAAC
MUT37
[0092] A Klebsiella bacterial mutant (MUT37) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding uridyltransferase (VIR37). The
insertion site nucleic acid sequence identifying the VIR37 gene in
MUT37 is shown in Table 39. TABLE-US-00057 TABLE 39 MUT37
Transposon Insertion Site (SEQ ID NO: 55)
CGAGCCACCCACTGTAGCGTATGGATATCGCGCAAGCCGCCGGGGCTGCTTTTCACGTCCGGCTCGA
GGTTATAGCTGGTGCCATGATAGCGCTGATGACGGACGTTCTGCTCTTCGACCTTGGCGGCGAAGAA
CTTTTCCGATGGCCAGAAGCCGTCGCTAAAAATATGTTTTTGCAGTTCAAGGAACAGCGCGACGTCG
CCGATCAGCAGGCGCGATTCGATTAAGTTGGTGGCAACGGTCAGATCCGAGAGACCTTCCAGCAGGC
ACTCTTCGAGGGTGCGTACGCTGTGGCCCACCTCCAGCTTGACGTCCCACAGCAGGGTGAGCAGTTC
GCCGACTTTTTGCGCCTGGTCGTCCGGCAGTTTTTTACGACTGAGGATCAGCAGATCGACGTCTGAG
AGCGGGTGCAG
MUT38
[0093] A Klebsiella bacterial mutant (MUT38) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding pyridoxine phosphate
biosynthetic protein PdxJ-PdxA (VIR38). The insertion site nucleic
acid sequence identifying the VIR38 gene in MUT38 is shown in Table
40. TABLE-US-00058 TABLE 40 MUT38 Transposon Insertion Site (SEQ ID
NO: 56)
CTTAACCCGCACGCTGGCGAAGGCGGCCATATGGGAACAGAAGAGATAGACACCATCATTCCGGTGC
TGGAAGAGATGCGCGCAAAGGGGATGAACCTCAGCGGTCCGCTGCCGGCAGACACTCTCTTTCAGCC
GAAATATCTTGATCATGCCGATGCGGTACTCGCGATGTACCACGATCAGGGCCTGCCCGTGCTAAAA
TACCAGGGCTTTGGCCGCCGCGTGAACATTACGCTCGGTTTACCTTTTATTCGTACCTCCGTCGACC
ACGGCACCGCACTGGAATTAGCGGGCCAGGGAAAAGCGGACGTCGGCAGTTTTATCACGGCGCTTAA
TCTCGCCATCAAAATGATTGTTAATACCCAATGAATAATCGAGTCCATCAGGGCCATTTAGCCCGCA
AACGCTTCGGGCAGAACTTCCTCAACGATCAGTTTGTGATCGACAGCATCGTCTCGGCGATTAACCC
GCAGAAAGGCCAGGCGATCGTTGAAATCGGC
MUT39
[0094] A Klebsiella bacterial mutant (MUT39) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding triose phosphate isomerase
(VIR39). The insertion site nucleic acid sequence identifying the
VIR39 gene in MUT39 is shown in Table 41. TABLE-US-00059 TABLE 41
MUT39 Transposon Insertion Site (SEQ ID NO: 57)
GGGTCTGACCCCGGTTCTGTGCATCGGTGAAACCGAAGCCGAAAACGAAGCGGGCAAAACGGAAGAA
GTTTGCGCACGTCAGATCGACGCCGTGCTGAAAACCCAGGGCGCTGCCGCTTTCGAAGGCGTGGTTA
TCGCTTACGAACCAGTATGGGCTATCGGTACCGGCAAATCAGCGACCCCGGCTCAGGCGCAGGCGGT
GCACAAATTCATCCGTGACCACATTGCTAAAGCTGACGCCAAAATCGCTGAGCAAGTGATCATCCAG
TACGGCGGTTCCGTTAACGCTGGCAACGCCGCAGAGCTGTTCACCCAGCCGGACATCGACGGCGCGC
TGGTTGGCGGCGCCTCCCTGAAAGCTGACGCTTTCGCGGTGATCGTTAAAGCAGCAGAAGCAGCGAA
AAAAGCGTAATTCGCTTTTCCCGGTGGCGACACGCGACCGGGTTGACTGACAAAACGTGGGAGCCCG
GCCT
MUT40
[0095] A Klebsiella bacterial mutant (MUT40) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding aldehyde dehydrogenase (VIR40).
The insertion site nucleic acid sequence identifying the VIR40 gene
in MUT40 is shown in Table 42. TABLE-US-00060 TABLE 42 MUT40
Transposon Insertion Site (SEQ ID NO: 58)
GGTGGCGCACCCTGGCGTCGTTTGTGTAGAAATTATGAATATTAATACCAGGAAAATTCCTAATTTT
TGTGTACGCTCTGACGAGCGCACAATAAAACAAGACGAATTTTTGAACAATTGTCTTTAAATTTGTT
AATTGAATTGATCTGTTGTTGTTTAAAGGTATTTGAATTTCTTTTGTATAGATATGTAAATTAACAT
TGAAAAGCCATTTCAAAAATTAAATATATGGCGAACATAGCTATTAACTTATAGTTAACATCTTCCC
GGGTTGCCTTTTGATACTTCGGGTAATATATTTATTTCGCACATCAAAATAACTCTTTTTTCTTCTG
TTTGTTATTCATGGCCATCTATTGGCGAAATAAGGCAGAGTAGAGGGGGATGTGCCTAATATCCTGC
GGAAGGAACGCAATGTACATTTACAGGGAGGAGCTGACGAGCCGTTTCGCGATAGCTTTAG
MUT41
[0096] A Klebsiella bacterial mutant (MUT41) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding galacosyl transferase (VIR41;
Clarke et al., J. Bacteriol., 177: 5411-18, 1995). The insertion
site nucleic acid sequence identifying the VIR41 gene in MUT41 is
shown in Table 43. TABLE-US-00061 TABLE 43 MUT41 Transposon
Insertion Site (SEQ ID NO: 59)
TTGGTGGTGTGCTCGCGAAGAAATTTAATCTGCCGGTCATCGTAAGTTTTGTTGGGCTTGGAAGAGT
ATTTTCTTCTGACAGCATGCCTTTAAAATTATTGCGGCAGTTTACTATTGCTGCATATAAATATATT
GCCAGTAATAAGCGCTGTATATTTATGTTTGAACATGACCGCGACAGAAAAAAACTGGCTAAGTTGG
TTGGACTCGAAGAACAACAGACTATTGTTATTGATGGTGCAGGCATTAATCCAGAGATATACAAATA
TTCTCTTGAACAGGATCACGATGTCCCTGTTGTATTGTTTGCCAGCCGTATGTTGTGGAGTAAAGGA
CTGGGCGACTTAATTGAAGCGAAGAAAATATTACGCAGTAAGAATATTCACTTTACTTTGAATGTTG
CTGGAATTCTGGTCGAAAATGATAAAGATGCAATTTCCCTTCAGGGTCATTGAAAATTGGCATCAGC
AAGGATTAATTAACTGGTTAGGTCGTTCGAATAATGTTTGCGATCTTATTGAGCAAT
MUT42
[0097] A Klebsiella bacterial mutant (MUT42) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding siroheme synthetase (VIR42;
Kolko et al., J. Bacteriol., 183: 328-35, 2001). The insertion site
nucleic acid sequence identifying the VIR42 gene in MUT42 is shown
in Table 44. TABLE-US-00062 TABLE 44 MUT42 Transposon Insertion
Site (SEQ ID NO: 60)
TTACTTGCCCCTTTTTGCCGAACTGAAACAAAGGCCCGTGCTGGTGATCGGCGGCGGCGAGATTGCT
GAACGTAAGATCAAGTTCCTGCTGCGCGCCCAGGCGCAGGTGCAGGTGGTCGCTGAAACGCTGTCAC
CGGCGCTGGCCGATCTGGCTGCGCGCCAGGCACTCAGCTGGCGGGCGACGGCATTCAGCGACTCGCT
GGTGGATGATGTCTTTCTGGTGATTGCGGCCACCGAGGATGAGGCGCTTAACCAGCGGGTGTTTGCG
GCAGCTAACGCGCGCTACCGGTTGGTCAACGTGGTGGATAACCAGGCGCTGTGCTCGTTTGTTTTCC
CTTCTATCGTCGACCGTTCGCCGCTGCTGGTGGCGATCTCCTCCAGCGGTAAAGCGCCGGTGTTGTC
GCGCATTCTGCGTGAAAAAATCGAAGCGCTGCTGCCGACGAATCTCGGTCGGCTGGCGCAATCAGCA
AGCT
MUT43
[0098] A Klebsiella bacterial mutant (MUT43) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding
7,8-dihydro-6-hydroxymethylpterin-pyrophosphokinase (VIR43). The
insertion site nucleic acid sequence identifying the VIR43 gene in
MUT43 is shown in Table 45. TABLE-US-00063 TABLE 45 MUT43
Transposon Insertion Site (SEQ ID NO: 61)
AGCAGGGCAATGGTGGTCGGTTTCATAACATTTCCTGATGATGAAAGTCATATTAACCGCCATTCTA
ACAGCAGCATTCAGAGGGGCAATGATTTTGGGCAACCGATTACGACGATCGCCGCAAATGCTAAAAA
AGGGAGAGGGGATTACCAGCTGGCGGGCTTTTCCGCGCCGAGATTATCCAGCACGGCGCGCAGCGCC
AGGCCGTCAGGAAAGTGAAGGTCCGGGGCGATCTCGAACAGCGGCCAGAGCATAAAGCCGCGGTTTT
TCATATCGTAGTGCGGAACGGTCAGGCGCTCGCTGTTAATGACAGCATCGCCAAACAGCATGATATC
GAGGTCCAGCGTGCGCGGCCCCCAGCGTTCGGCTTTGCGCACTCGCCCCTGCTGCAGTTCGATGCGC
TGAGTATGATCGAGCAGCGTCTCGGGGGGCAGGGCGGTTTCCAGCGCAA
MUT44
[0099] A Klebsiella bacterial mutant (MUT44) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding glucose-6-phosphate isomerase
(VIR44). The insertion site nucleic acid sequence identifying the
VIR44 gene in MUT44 is shown in Table 46. TABLE-US-00064 TABLE 46
MUT44 Transposon Insertion Site (SEQ ID NO: 62)
GGCTTAACGCCAGCTATGTCAACGCTGCGGTTATGCGGATTTTTCATGCCTCTGCGGCTAACAGAAA
AAAGCCTTATGATAGCTATACTAATGGGGCTTTTTACTCCGTTTTGACCCGATTCCTGACCGGCGTC
AGGGTCAAGTCACAAAAATCATCACAATTTTCCGTCACCGGCGCTACAATCGACCGAAGTCACAATC
TCAAATCAGAAGAGTATTGCTAATGAAAAACATCAACCCAACGCAGACCTCTGCCTGGCAGGCATTA
CAGAAACACTTCGACGAAATGAAAGATGTCACTATCAGCGAGCTTTTCGCCAAAGATAGCGACCGTT
TTTCTAAATTTTCCGCGACGTTCGACGATCTGATGCTGGTGGACTTCTCCAAAAACCGCATCACTGA
AGAGACGCTGGCTAAACTGCAGGATCTGGCGAAAGAGACTGACCTGGCGGGCGCTATCAAGTCGATG
TTCTCAGGTGAGAAGATCAACCGCACCGAAGACCGCGCGGTACTGCACGTCGCGCT
MUT45
[0100] A Klebsiella bacterial mutant (MUT45) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding DNA methylase (VIR45). The
insertion site nucleic acid sequence identifying the VIR45 gene in
MUT45 is shown in Table 47. TABLE-US-00065 TABLE 47 MUT45
Transposon Insertion Site (SEQ ID NO: 63)
TGCTTCATCCGCATCTCCTTGAAATTTATTTGGTCTTAGGCGGACGGTAGAGCGCTAATAGCTCGTC
CACCTTTTTACGCGTACCACCGTTGCTGCTGATGCTGCGCCGCACCTTCACAATATGCGTTTCTGCC
GCGTTTTTATACCATTCCTGCGTCAGCGGCGTGCGGTGGTTGGAAATCAGCACCGGGATGCGCTTTT
TCATCAGCGATTCCGCCTTTTGCGCCAGCAGTACCTGTTGTTCCAGGTTGAAACTGTTGGTGTGGTA
GGCGGTAAAGTTCGCCGTCGCCGTTAGCGGCGCATAGGGCGGATCGCAATACACCACTGTGCGGCTA
TCCGCACGTTGCATGCACTCTTCGTAAGATTCGCAGTAAAACTCGGCGTTTTGCGCCTTCTCGGCGA
AATGATAGAGCTCAGCTTCGGGGAAATAGGGCTTTTTATAACGGCCAAACGGCACATTGAACTCGCC
GCGCAG
MUT46
[0101] A Klebsiella bacterial mutant (MUT46) was made by transposon
insertion in a Klebsiella sp. wild-type strain. In the
Dictyostelium growth assay, the mutated microorganism was less
virulent compared to an isogenic bacterial strain. The nucleotide
sequence immediately following the transposon insertion was cloned
and identified as the gene encoding a putative inner membrane
protein (VIR46). The insertion site nucleic acid sequence
identifying the VIR46 gene in MUT46 is shown in Table 48.
TABLE-US-00066 TABLE 48 MUT46 Transposon Insertion Site (SEQ ID NO:
64)
TGTCAATGCGCAATTTGGTTAAATATGTCGGTATTGGCCTGCTGGTGATGGGGCTTGCCGCCTGCGA
TAACAGCGATTCAAAAGCGCCAACCGTTGGCGCAGCAGCGGAGAGCAATGCCAGCGGCCAGGCAATC
AGCCTGCTGGATGGCAAGCTGAGCTTCACCCTGCCTGCGGGCATGGCCGACCAGAGCGGCAAACTGG
GTACCCAGGCGAACAATATGCACGTCTACTCTGACGCTACCGGCCAGAAAGCGGTCATCGTCATCGT
CGGCGACAGCACCAATGA
IV. Suitable Target Pathogens
[0102] Other Pseudomonas sp. and Klebsiella sp. and many other
microbes, including gram-negative bacterial strains, are likely to
include virulence genes encoding VIRX-related peptides or proteins
having amino acid sequence identity or similarity to those
identified herein. Suitable bacterial pathogens may include, but
are not limited to, Pneumococci sp., Klebsiella, sp., Pseudomonas,
e.g., P. aeruginosa, Salmonella, e.g., Salmonella typhimurium,
Legionella, e.g., Legionella pneumophilia, Escherichia, e.g.,
Escherichia coli, Listeria, e.g., Listeria monocytogenes,
Staphylococcus, e.g., Staphylococcus aureus, Streptococci sp.,
Vibrio, e.g., Vibrio cholerae. Pathogenic mycobacteria of the
present invention may include e.g., Mycobacterium tuberculosis.
Pathogenic fungi of the present invention may include, e.g.,
Candida albicans. Pathogenic unicellular eukaryotic organisms of
the present invention may include, e.g., Leishmania donovani.
[0103] Having identified VIRX genes according to the invention, it
is possible to use the gene sequence to search for related genes or
peptides in other microorganisms. This may be carried out by
searching in existing databases, e.g., EMBL or GenBank. The levels
of identity between gene sequences and levels of identity or
similarity between, amino acid sequences can be calculated using
known methods. In relation to the present invention, publicly
available computer based methods for determining identity and
similarity include the BLASTP, BLASTN and FASTA (Atschul et al., J.
Molec. Biol., 1990; 215:403-410), the BLASTX program available from
NCBI, and the Gap program from Genetics Computer Group, Madison
Wis.
[0104] Preferably, the peptides that may be useful in the various
aspects of the invention have greater than a 40% similarity with
the peptides identified herein. More preferably, the peptides have
greater than 60% sequence similarity. Most preferably, the peptides
have greater than 80% sequence similarity, e.g., 95% similarity.
With regard to the polynucleotide sequences identified herein,
related polynucleotides that may be useful in the various aspects
of the invention may have greater than 40% identity with the
sequences identified herein. More preferably, the polynucleotide
sequences have greater than 60% sequence identity. Most preferably,
the polynucleotide sequences have greater than 80% sequence
identity, e.g., 95% identity.
[0105] In addition to related molecules from other microorganisms,
the invention encompasses modifications made to the peptides and
polynucleotides identified herein which do not significantly alter
the biological function. It will be apparent to the artisan that
the degeneracy of the genetic code can result in polynucleotides
with minor base changes from those specified herein, but which
nevertheless encode the same peptides. Complementary
polynucleotides are also within the invention. Conservative
replacements at the amino acid level are also envisaged, i.e.,
different acidic or basic amino acids may be substituted without
substantial loss of function.
[0106] It is recognized in the art that highly refined mechanisms
that regulate transcription have evolved and are present in
bacteria. Most bacterial genes are organized into operons, which
are groups of genes coding for related proteins. Operons can either
be repressed or induced thus regulating those genes. An operon
consists of an operator, promoter, regulator, and structural genes.
The regulator gene codes for a repressor protein that binds to the
operator, obstructing the promoter (thus, transcription) of the
structural genes. The regulator does not have to be adjacent to
other genes in the operon. If the repressor protein is removed,
transcription may occur.
[0107] Transposon mutagenesis usually inactivates the gene in which
the transposon is inserted, as well as any gene downstream in the
same operon. If the VIRX gene is a structural gene in an operon,
inactivation of the MAX gene disrupts the expression of other
structural genes in the same operon and positioned downstream of
the inactivated VIRX gene. For example, an insertion in pchE gene
also inactivates pchF, pchG, pchH, and pchI genes because they all
reside within the pchEFGHI operon and are downstream of the
inactivated pchE gene. Accordingly, the present invention includes
attenuation of virulence due to alteration of a VIRX gene residing
in an operon as well as alterations to nucleic acid yielding loss
of expression of structural genes located in the same operon and
located downstream of the VIRX gene. In one embodiment, the present
invention is an alteration inactivating the first gene of an operon
carrying a VIRX gene of the invention. The alteration of nucleic
acids of VIRX genes and VIRX-containing operons may be insertional
inactivation or gene deletion. It is preferred that the alteration
of nucleic acids of VIRX genes and VIRX-containing operons be
insertional inactivation.
[0108] The present invention also provides for a bacterial strain
comprising an operon encoding a gene selected from the group
consisting of VIR1, VIR2, VIR3, VIR4. VIR5, VIR6, VIR7, VIR8, VIR9,
VIR10, VIR11, VIR12, VIR13, VIR14, VIR15, VIR16, VIR17, VIR18,
VIR19, VIR20, VIR21, VIR22, VIR23, VIR24, VIR25, VIR26, VIR27,
VIR28, VIR29, VIR30, VIR31, VIR32, VIR33, VIR34, VIR35, VIR36,
VIR37, VIR38, VIR39, VIR40, VIR41, VIR42, VIR44, VIR45, and VIR46,
wherein the bacterial strain includes a mutation that reduces
expression of the VIRX gene relative to an isogenic bacterial
strain lacking the mutation. In one embodiment, the mutation
reduces inhibition of Dictyostelium amoeba growth when compared to
the growth of Dictyostelium amoeba in the presence of an isogenic
bacterial strain lacking the mutation. In another embodiment, the
attenuated bacterial strain has more than one mutation of an operon
containing a VIRX gene when compared to an isogenic bacterial
strain.
V. VIRX Nucleic Acids and Polypeptides Can Be Used to Identify
Antimicrobial Drugs
[0109] A. Screening
[0110] In a separate embodiment, the VIRX genes, or their
polynucleotide or polypeptide products disclosed herein is used in
screening assays for the identification of potential antimicrobial
drugs. Routine screening assays are known to those skilled in the
art, and can be adapted using the VIRX products of the invention in
the appropriate way. For example, the products of the invention can
be used as the target for a potential drug, with the ability of the
drug to inactivate or bind to the tar-get indicating its potential
antimicrobial activity. In the methods of the present invention,
one or more test compounds may be present or produced in the assay
mixture. Preferably one compound is present, or produced, in the
assay mixture.
[0111] B. Character of Antimicrobial Candidate Compositions
[0112] VIRX nucleic acids and polypeptides may be used to identify
drugs or therapeutics in a candidate composition useful in the
prevention or treatment of pathogen-associated disease or
infection. A candidate composition can include one or more
molecules for analysis in a screening assay and can be a synthetic
or semi-synthetic molecules. Such molecules include inorganic as
well as organic chemical molecules. The molecules may be less than
about 500 Daltons or more than 500 Daltons. The molecules may be
naturally occurring. Naturally occurring molecules may include,
e.g., saccharides, lipids, peptides, proteins, nucleic acids, or
combinations thereof, e.g., aminoglycosides, glycolipids,
lipopolysaccharides, or macrolides. Proteins may be
immunoglobulins, e.g., polyclonal or monoclonal antibodies. Nucleic
acids may be DNA or RNA, e.g., small interfering RNA (siRNA). The
precise source of the molecule is not critical to the method of the
present invention. The molecule might be derived from e.g.,
synthetic compounds libraries that are commercially available,
e.g., Sigma-Aldrich (Milwaukee, Wis.), or libraries of natural
occurring molecules in the form of bacterial, fungal, plant, and
animal extracts such as those available from Xenova (Slough, UK).
The synthetic (or semi-synthetic) or natural occurring molecules
might be modified using standard chemical, physical, or biochemical
methods known in the art.
VI. VIRX Nucleic Acids and Polypeptides Can Be Used to Detect the
Degree of Virulence of Pathogens
[0113] A diagnostic test can assist physicians in determining the
type of disease and appropriate associated therapy. As such, a
separate embodiment of this invention provides for the use of VIRX
genes or their polynucleotides or nucleic acid products as
virulence markers for detecting the presence of a pathogen, a
pathogen-associated disease, or the virulence of a pathogen. There
are many diagnostic assay approaches known to the artisan.
Generally, the diagnostic method used would comprise the steps of
(a) obtaining a sample from a potentially diseased subject or a
diseased subject; (b) measuring the level of at least one
polypeptide or polynucleotide virulence marker in the sample; and
(c) comparing the amount of the virulence marker in the sample of
step (a) to the amount of the virulence marker present in a control
sample from a second subject known not to have the presence of the
pathogen, where an alteration in the expression level of the
virulence marker in the first subject as compared to the control
sample indicates the presence of a pathogen, a pathogen-associated
disease, or the virulence of a pathogen. Preferably, the subject is
a mammal. More preferred is that the subject is a human. The person
of skill will recognize that diagnostic tests may be performed in
an array-type format wherein, e.g., the presence of two or more
VIRX genes or gene products indicate the presence of a pathogen, a
pathogen-associated disease, or the virulence of a pathogen.
VII. Attenuated Organisms of the Present Invention Can Be Used in
Vaccine Preparation
[0114] In another embodiment, the invention provides for the use of
the attenuated organisms described herein in vaccine preparation.
The preparation of vaccines based on attenuated microorganisms is
known to those skilled in the art. Vaccine compositions can be
formulated with suitable carriers or adjuvants, e.g., alum, as
necessary or desired, to provide effective immunization against
infection. The preparation of vaccine formulations will be apparent
to the artisan. The attenuated microorganisms may be prepared with
a mutation that disrupts the expression of any of the VIRX genes
identified herein. The artisan will be aware of methods for
disrupting expression of particular VIRX genes. Techniques that may
be used include, but are not limited to, insertional inactivation,
or gene deletion techniques. Attenuated microorganisms according to
the invention may also comprise additional mutations in other
genes, for example in a second gene identified herein or in a
separate gene required for growth of the microorganism, e.g., an
Aro mutation. Attenuated microorganisms may also be used as carrier
systems for the delivery of heterologous antigens, therapeutic
proteins or nucleic acids (DNA or RNA). In this embodiment, the
attenuated microorganisms are used to deliver a heterologous
antigen, protein or nucleic acid to a particular site in vivo.
Introduction of a heterologous antigen, peptide or nucleic acid
into an attenuated microorganism can be carried out by conventional
techniques, including the use of recombinant constructs, e.g.,
vectors, which comprise polynucleotides that express the
heterologous antigen or therapeutic protein, and also include
suitable promoter sequences. Alternatively, the gene that encodes
the heterologous antigen or protein may be incorporated into the
genome of the organism and the endogenous promoters used to control
expression. In the vaccines of the present invention, the
pharmaceutically effective dosage of the mutants of the present
invention to be administered may vary depending on the age, weight
and sex of the subject, and the mode of administration. The subject
can be, e.g., a human, a non-human primate (such as an ape,
gorilla, or chimpanzee), cow, horse, pig, sheep, dog, cat, or
rodent (including mouse or rat).
VIII. Definitions
[0115] As used herein, each of the following terms has the meaning
associated with it in this section.
[0116] The term "pathogen," as used herein, is intended to include
an agent that causes disease, especially a living microorganism
such as a bacterium or fungus. The terms "agent" and "factor" are
used interchangeably herein to describe pathogens or toxins useful
in the methods of the present invention. Pathogens may include any
bacteria, mycobacteria, fungi and unicellular eukaryotic organism,
including wild types and mutants thereof, which causes disease or
brings about damage or harm to a host organism. Pathogens may also
be a poisonous substance, e.g., toxin, which is produced by living
cells or organisms and is capable of causing disease when
introduced to a host.
[0117] The term, "pathogenic," as used herein, is defined as an
agent's ability to cause disease, damage or harm to a host
organism.
[0118] The term, "attenuated," as used herein, means an organism
made less virulent relative to an isogenic pathogenic organism.
[0119] The term, "virulence," as used herein, is a measure of the
degree of pathogenicity of an agent to a host organism. Virulence
is usually expressed as the dose of an agent or cell number of a
pathogen that will elicit a pathological response in the host
organism within a given time period. "Reducing the virulence" as
used herein is defined as the ability of a compound to attenuate,
diminish, decrease, suppress, or arrest the development of, or the
progression of disease, damage or harm to a host organism mediated
by a pathogen.
[0120] The term, "host organism," as used herein, is intended to
include any living organism. Preferably the host organism is a
eukaryote, e.g., vertebrate. More preferably the host organism is a
mammal. It is most preferred that the host organism be a human.
[0121] The term, "mutant," as used herein, an organism carrying a
specific mutation of a gene that is expressed in the organism's
phenotype.
[0122] The term, "mutation," as used herein, is an alteration of
one or more nucleic acids of a polynucleotide sequence encoding a
gene. A mutation may include the insertion of additional nucleic
acids to a polynucleotide sequence encoding a gene, e.g.,
insertional inactivation of a gene. Alternatively, a mutation may
include, but is not limited to, deletion of one or more nulceic
acids of a polynucleotide sequence encoding a gene.
[0123] The term, "operon," as used herein, is a unit of bacterial
gene expression and regulation comprising several genes usually
with complementary functions. Typically an operon includes nucleic
acid and control elements in the nucleic acid that may be
recognized by regulators of gene products. Insertion in a gene in
an operon interferes with the function of this gene and of other
genes located downstream or upstream in the operon. It is
understood herein that the function attributed to a gene refers to
its function and/or that of any gene located downstream or upstream
in the same operon.
[0124] The term, "pharmaceutically effective dosage," as used
herein, means that amount necessary at least partly to attain the
desired effect, or to delay the onset of, inhibit the progression
of, or halt altogether, the onset or progression of the particular
condition being treated.
[0125] The terms "similarity" and "identity" are known in the art.
The use of the term "identity" refers to a sequence comparison
based on identical matches between correspondingly identical
positions in the sequences being compared. The term "similarity"
refers to a comparison between amino acid sequences, and takes into
account not only identical amino acids in corresponding positions,
but also functionally similar amino acids in corresponding
positions. Thus similarity between polypeptide sequences indicates
functional similarity, in addition to sequence similarity.
EQUIVALENTS
[0126] From the foregoing detailed description of the specific
embodiments of the invention, it should be apparent that bacterial
genes have been identified and assigned a new role in virulence.
Further, these genes and their products are useful in the
identification of antimicrobial agents, the diagnosis of
pathogen-associated disease or infection as well as the preparation
of vaccines. Although particular embodiments have been disclosed
herein in detail, this has been done by way of example for purposes
of illustration only, and is not intended to be limiting with
respect to the scope of the appended claims that follow. In
particular, it is contemplated by the inventor that various
substitutions, alterations, and modifications may be made to the
invention without departing from the spirit and scope of the
invention as defined by the claims. For instance, the choice of the
particular pathogen, or combination of pathogens selected for assay
or vaccination, the test conditions used in diagnostic assays
utilizing the pathogens of this invention, or the method of
mutagenesis used to derive the attenuated mutants is believed to be
a matter of routine for a person of ordinary skill in the art with
knowledge of the embodiments described herein.
EXAMPLES
[0127] This Example is provided for the purpose of illustration
only and the invention should in no way be construed as being
limited to these Example, but rather should be construed to
encompass any and all variations which become evident as a result
of the teaching provided.
Example 1
Strains and Culture Conditions Used to Screen for Attenuated
Viurlence in Test Bacterial Mutants
[0128] The D. discoideum wild-type strain DH1-10 used in these
studies is a subclone of DH1 (Cornillon et al., J. Biol. Chem.,
275(44):34287-92, 2000). Cells were grown at 21.degree. C. in HL5
medium (14.3 g/l peptone (Oxoid), 7.15 g/l yeast extract, 18 g/l
maltose, 0.64 g/l Na.sub.2HPO.sub.4.2H.sub.2O, 0.49 g/l
KH.sub.2PO.sub.4, pH 6.7) (Cornillon et al., J. Cell. Sci., 107 (Pt
10):2691-704, 1994) and subcultured twice a week.
[0129] Bacteria were grown overnight at 37.degree. C. on
Luria-Bertani (LB) agar. Single colonies were inoculated into 5 ml
PB (2% (wt/vol) peptone, 0.3% (wt/vol) MgCl.sub.2.6H.sub.2O, 1%
(wt/vol) K.sub.2SO.sub.4) (Essar et al., J. Bacteriol.,
172(2):884-900,1990) in a 50 ml flask and grown at 37.degree. C.
for 8 hr prior to use. The growth of various strains was tested in
rich medium (PB) by measuring the optical density (600 nm) of a
culture at different times after inoculation and was found to be
comparable for all strains used. Under these conditions, similar
OD.sub.600s were obtained for each strain and the induction of
quorum sensing was maximal. Minimal Inhibitory Concentrations
(MICs) were determined in Mueller-Hinton broth by the microdilution
method (Thornsberry et al., NCCLS, 3: 48-56, 1983). Mutations
yielding reduced virulence were identified where the growth of the
Dictyostelium test host organism exposed to the mutant pathogen was
greater than the Dictyostelium test host organism exposed to
wild-type pathogen. Specific genetic mutations in pathogens
displaying reduced virulence were identified and characterized by
techniques well know in the art.
Sequence CWU 1
1
64 1 1050 DNA Pseudomonas aeruginosa 1 atggatatca agggagccct
caatcgcatc gtcaaccagc tcgacctgac caccgaggaa 60 atgcaggcgg
tcatgcgcca gatcatgacc gggcagtgca ccgacgcgca gatcggcgcc 120
ttcctgatgg gcatgcggat gaagagcgaa accatcgacg agatcgtcgg cgcggtggcg
180 gtgatgcgcg aactggccga cggcgtgcag ttgcctacgc tgaagcatgt
ggtcgacgtg 240 gtcggcaccg gcggcgatgg cgcgaacatc ttcaacgtgt
cctcggcggc gtccttcgtg 300 gtcgccgccg ctggcggcaa ggtcgccaaa
cacggtaacc gcgcggtctc cggcaagagc 360 ggcagcgccg acttgctgga
agccgccggc atctacctgg agctgacctc cgaacaggtg 420 gcgcgttgca
tcgacaccgt cggcgtcggg ttcatgttcg cccaggtcca ccacaaggcg 480
atgaagtacg ccgccggtcc gcgccgcgag ctgggcttgc ggactctgtt caacatgctt
540 ggcccactga ccaacccggc gggagtcagg caccaggtgg tcggggtgtt
cacccaggaa 600 ctgtgcaagc cgctggctga agtgctcaag cgtctcggca
gcgagcatgt gctggtggtg 660 cattcgcgcg acgggctgga cgagttcagt
ctggccgcgg cgacccacat tgccgagttg 720 aaggacggcg aggtacgcga
gtacgaagtg cgtcccgagg acttcgggat caagagccag 780 accctgatgg
ggctggaggt cgacagtccg caggcctcgc tggaactgat ccgcgacgct 840
ttggggcggc gcaagaccga ggctgggcag aaggccgccg agctgatcgt gatgaatgcc
900 ggcccggcac tgtacgctgc cgatctggcg accagcctgc acgagggcat
tcaactggcc 960 cacgatgccc tgcacaccgg gctggcacgg gagaagatgg
acgaactggt ggccttcacc 1020 gccgtttaca gagaggagaa cgcacagtga 1050 2
349 PRT Pseudomonas aeruginosa 2 Met Asp Ile Lys Gly Ala Leu Asn
Arg Ile Val Asn Gln Leu Asp Leu 1 5 10 15 Thr Thr Glu Glu Met Gln
Ala Val Met Arg Gln Ile Met Thr Gly Gln 20 25 30 Cys Thr Asp Ala
Gln Ile Gly Ala Phe Leu Met Gly Met Arg Met Lys 35 40 45 Ser Glu
Thr Ile Asp Glu Ile Val Gly Ala Val Ala Val Met Arg Glu 50 55 60
Leu Ala Asp Gly Val Gln Leu Pro Thr Leu Lys His Val Val Asp Val 65
70 75 80 Val Gly Thr Gly Gly Asp Gly Ala Asn Ile Phe Asn Val Ser
Ser Ala 85 90 95 Ala Ser Phe Val Val Ala Ala Ala Gly Gly Lys Val
Ala Lys His Gly 100 105 110 Asn Arg Ala Val Ser Gly Lys Ser Gly Ser
Ala Asp Leu Leu Glu Ala 115 120 125 Ala Gly Ile Tyr Leu Glu Leu Thr
Ser Glu Gln Val Ala Arg Cys Ile 130 135 140 Asp Thr Val Gly Val Gly
Phe Met Phe Ala Gln Val His His Lys Ala 145 150 155 160 Met Lys Tyr
Ala Ala Gly Pro Arg Arg Glu Leu Gly Leu Arg Thr Leu 165 170 175 Phe
Asn Met Leu Gly Pro Leu Thr Asn Pro Ala Gly Val Arg His Gln 180 185
190 Val Val Gly Val Phe Thr Gln Glu Leu Cys Lys Pro Leu Ala Glu Val
195 200 205 Leu Lys Arg Leu Gly Ser Glu His Val Leu Val Val His Ser
Arg Asp 210 215 220 Gly Leu Asp Glu Phe Ser Leu Ala Ala Ala Thr His
Ile Ala Glu Leu 225 230 235 240 Lys Asp Gly Glu Val Arg Glu Tyr Glu
Val Arg Pro Glu Asp Phe Gly 245 250 255 Ile Lys Ser Gln Thr Leu Met
Gly Leu Glu Val Asp Ser Pro Gln Ala 260 265 270 Ser Leu Glu Leu Ile
Arg Asp Ala Leu Gly Arg Arg Lys Thr Glu Ala 275 280 285 Gly Gln Lys
Ala Ala Glu Leu Ile Val Met Asn Ala Gly Pro Ala Leu 290 295 300 Tyr
Ala Ala Asp Leu Ala Thr Ser Leu His Glu Gly Ile Gln Leu Ala 305 310
315 320 His Asp Ala Leu His Thr Gly Leu Ala Arg Glu Lys Met Asp Glu
Leu 325 330 335 Val Ala Phe Thr Ala Val Tyr Arg Glu Glu Asn Ala Gln
340 345 3 918 DNA Pseudomonas aeruginosa 3 atggtcgaca aactgacgca
cctgaaacag ctggaggcgg aaagcatcca catcatccgc 60 gaggtggccg
ccgagttcga taacccggtg atgctgtact cgatcggcaa ggattccgcg 120
gtcatgctgc acctggcccg caaggccttc ttccccggca agctgccctt cccggtgatg
180 cacgtggaca cccgctggaa attccaggag atgtacaggt tccgtgatcg
gatggtcgag 240 gaaatgggcc tggatctgat cacccacgtc aacccggacg
gcgtcgccca gggcatcaac 300 ccgttcaccc acggcagcgc caagcacacc
gacgtgatga agaccgaggg actcaagcag 360 gccctggaca agtacggttt
cgacgctgcc ttcggcggtg cgcgccgcga cgaggagaag 420 tcgcgggcca
aggaacgggt ctattcgttc cgcgacagca agcaccgctg ggacccgaag 480
aaccagcgtc ccgagctgtg gaacatctac aacggcaagg tgaagaaggg cgagtcgatc
540 cgcgtcttcc cgctgtccaa ctggaccgag ctggacatct ggcaatacat
ctacctggaa 600 ggcatcccga tcgtcccgct gtacttcgcc gccgagcgcg
aggtcatcga gaagaatggc 660 acattgatca tgatcgacga cgagcgcatc
ctcgagcatc tctctgacga agagaaagcc 720 cgcatcgaga agcgcatggt
gcgcttccgt accctcggct gctacccgct caccggcgcg 780 gtcgagtcca
gcgccaccac gctgccggaa atcatccagg aaatgctcct gacgcgtact 840
tccgaacgcc agggccgggt catcgaccat gaccaggccg gttcgatgga agaaaagaaa
900 cgtcagggct atttctga 918 4 305 PRT Pseudomonas aeruginosa 4 Met
Val Asp Lys Leu Thr His Leu Lys Gln Leu Glu Ala Glu Ser Ile 1 5 10
15 His Ile Ile Arg Glu Val Ala Ala Glu Phe Asp Asn Pro Val Met Leu
20 25 30 Tyr Ser Ile Gly Lys Asp Ser Ala Val Met Leu His Leu Ala
Arg Lys 35 40 45 Ala Phe Phe Pro Gly Lys Leu Pro Phe Pro Val Met
His Val Asp Thr 50 55 60 Arg Trp Lys Phe Gln Glu Met Tyr Arg Phe
Arg Asp Arg Met Val Glu 65 70 75 80 Glu Met Gly Leu Asp Leu Ile Thr
His Val Asn Pro Asp Gly Val Ala 85 90 95 Gln Gly Ile Asn Pro Phe
Thr His Gly Ser Ala Lys His Thr Asp Val 100 105 110 Met Lys Thr Glu
Gly Leu Lys Gln Ala Leu Asp Lys Tyr Gly Phe Asp 115 120 125 Ala Ala
Phe Gly Gly Ala Arg Arg Asp Glu Glu Lys Ser Arg Ala Lys 130 135 140
Glu Arg Val Tyr Ser Phe Arg Asp Ser Lys His Arg Trp Asp Pro Lys 145
150 155 160 Asn Gln Arg Pro Glu Leu Trp Asn Ile Tyr Asn Gly Lys Val
Lys Lys 165 170 175 Gly Glu Ser Ile Arg Val Phe Pro Leu Ser Asn Trp
Thr Glu Leu Asp 180 185 190 Ile Trp Gln Tyr Ile Tyr Leu Glu Gly Ile
Pro Ile Val Pro Leu Tyr 195 200 205 Phe Ala Ala Glu Arg Glu Val Ile
Glu Lys Asn Gly Thr Leu Ile Met 210 215 220 Ile Asp Asp Glu Arg Ile
Leu Glu His Leu Ser Asp Glu Glu Lys Ala 225 230 235 240 Arg Ile Glu
Lys Arg Met Val Arg Phe Arg Thr Leu Gly Cys Tyr Pro 245 250 255 Leu
Thr Gly Ala Val Glu Ser Ser Ala Thr Thr Leu Pro Glu Ile Ile 260 265
270 Gln Glu Met Leu Leu Thr Arg Thr Ser Glu Arg Gln Gly Arg Val Ile
275 280 285 Asp His Asp Gln Ala Gly Ser Met Glu Glu Lys Lys Arg Gln
Gly Tyr 290 295 300 Phe 305 5 822 DNA Pseudomonas aeruginosa 5
atgaggccgg tgccttgggg cgaattggtg gcgctggtgc ggcgcgccgg cgaggcgatc
60 ctgccgcact ggcgcgccga cgtggtggtg cgctcgaagg ccgacgaatc
gccggtgact 120 gccgccgacc tggccgcgca ccatatattg gaggcgggat
tgcgggcgct ggcgccggac 180 attccggtgc tttccgaaga ggattgcgag
ataccgctga gcgagcgcgg ccactggcgg 240 cgctggtggc tggtggaccc
gctggacggc accaaggagt tcatctccgg tagcgaggag 300 ttcaccgtca
acgtggccct ggtcgaggat ggccgggtgc tgttcggcct ggtcggcgtg 360
ccggtgagcg gccgctgcta ctacggtggc gccggtctcg gtgcctggcg cgaggaggcc
420 gatggccgcg cgcaaccgat cagtgtgcgc ctggagcccg aggaggcctt
caccgtggtg 480 gccagcaagc gccatggcag cccggcccag gagcgcctgc
tggatggctt gagcgagcgc 540 ttcggcgacc tgcggcgagc cagcatcggc
agttcgctga agttctgcct gctggccgag 600 ggcgctgccg actgctatcc
gcgcctgacg ccaacctcgc aatgggacac ggccgccgcc 660 cagggtgtgc
tggaaggcgc cggcggcgag gtgctcgacc tgcatggtgc gccattcacc 720
tacgagccgc gcgaggatta cctcaacggc tccttcctgg ccctgccgcg cgccgccgag
780 tggcgcagcg agctgatcca actggcgcgc gcgctgcact ga 822 6 273 PRT
Pseudomonas aeruginosa 6 Met Arg Pro Val Pro Trp Gly Glu Leu Val
Ala Leu Val Arg Arg Ala 1 5 10 15 Gly Glu Ala Ile Leu Pro His Trp
Arg Ala Asp Val Val Val Arg Ser 20 25 30 Lys Ala Asp Glu Ser Pro
Val Thr Ala Ala Asp Leu Ala Ala His His 35 40 45 Ile Leu Glu Ala
Gly Leu Arg Ala Leu Ala Pro Asp Ile Pro Val Leu 50 55 60 Ser Glu
Glu Asp Cys Glu Ile Pro Leu Ser Glu Arg Gly His Trp Arg 65 70 75 80
Arg Trp Trp Leu Val Asp Pro Leu Asp Gly Thr Lys Glu Phe Ile Ser 85
90 95 Gly Ser Glu Glu Phe Thr Val Asn Val Ala Leu Val Glu Asp Gly
Arg 100 105 110 Val Leu Phe Gly Leu Val Gly Val Pro Val Ser Gly Arg
Cys Tyr Tyr 115 120 125 Gly Gly Ala Gly Leu Gly Ala Trp Arg Glu Glu
Ala Asp Gly Arg Ala 130 135 140 Gln Pro Ile Ser Val Arg Leu Glu Pro
Glu Glu Ala Phe Thr Val Val 145 150 155 160 Ala Ser Lys Arg His Gly
Ser Pro Ala Gln Glu Arg Leu Leu Asp Gly 165 170 175 Leu Ser Glu Arg
Phe Gly Asp Leu Arg Arg Ala Ser Ile Gly Ser Ser 180 185 190 Leu Lys
Phe Cys Leu Leu Ala Glu Gly Ala Ala Asp Cys Tyr Pro Arg 195 200 205
Leu Thr Pro Thr Ser Gln Trp Asp Thr Ala Ala Ala Gln Gly Val Leu 210
215 220 Glu Gly Ala Gly Gly Glu Val Leu Asp Leu His Gly Ala Pro Phe
Thr 225 230 235 240 Tyr Glu Pro Arg Glu Asp Tyr Leu Asn Gly Ser Phe
Leu Ala Leu Pro 245 250 255 Arg Ala Ala Glu Trp Arg Ser Glu Leu Ile
Gln Leu Ala Arg Ala Leu 260 265 270 His 7 1299 DNA Pseudomonas
aeruginosa 7 atgcgagttc tggtccttgg cagcggtgtc atcggtaccg ccagtgcgta
ttacctggcc 60 cgtgccgggt tcgaggtggt ggtggtcgac cgtcaggacg
gtcccgcgct ggaaaccagc 120 ttcgccaacg ccggccaggt gtctcccggc
tacgcttcgc cctgggcagc cccgggcatt 180 cccctgaagg ccatgaagtg
gctgctggaa aagcacgcgc cgctggccat caagctcacc 240 tccgatccca
gccagtacgc ctggatgctg cagatgctgc gcaactgcac cgccgagcgc 300
tacgccgtga acaaggagcg catggtccgc ctgtccgagt acagccgcga ttgcctcgac
360 gaactgcgcg ccgagaccgg catcgcctac gagggccgca ccctcggcac
cacccaactg 420 ttccgcaccc aggcgcagct ggacgccgcc ggcaaggaca
tcgccgtgct cgagcgctcc 480 ggcgtgccct acgaggttct cgaccgcgac
ggcatcgccc gcgtagagcc ggctttggcc 540 aaggtcgccg acaagctggt
cggcgccttg cgcctgccca acgaccagac cggcgactgc 600 cagctgttca
ccacccgcct ggcggaaatg gccaagggcc tgggcgtgga gttccgcttc 660
ggccagaaca tcgagcgcct ggacttcgcc ggcgaccgca tcaacggcgt gctggtcaac
720 ggcgaattgc tcaccgccga ccactacgtg ctggccctgg gcagctactc
gccgcaactg 780 ctcaagccgc tgggtatcaa ggctccggtc tatccgctga
agggttattc gctgaccgtg 840 ccgatcacca acccggagat ggcgccgacc
tcgaccatcc tcgacgagac ctacaaggtg 900 gcgatcaccc gcttcgacca
gcgcatccgc gtcggcggca tggcggaaat cgccggcttc 960 gacctgtcgc
tgaacccgcg ccgccgcgag accctggaaa tgatcaccac cgacctctat 1020
cccgagggcg gcgatatcag ccaggcgacc ttctggaccg gcctgcgccc ggcgaccccg
1080 gatggcaccc cgatcgtcgg cgccacccgc taccgcaacc tgttcctcaa
taccggccac 1140 ggcaccctgg gttggaccat ggcctgcggg tcgggtcgct
acctggccga cctgatggcg 1200 aagaagcgcc cgcagatcag taccgaaggc
ctggatattt cccgctacag caattccccg 1260 gagaacgcca agaatgccca
tccagcgcca gcacactaa 1299 8 432 PRT Pseudomonas aeruginosa 8 Met
Arg Val Leu Val Leu Gly Ser Gly Val Ile Gly Thr Ala Ser Ala 1 5 10
15 Tyr Tyr Leu Ala Arg Ala Gly Phe Glu Val Val Val Val Asp Arg Gln
20 25 30 Asp Gly Pro Ala Leu Glu Thr Ser Phe Ala Asn Ala Gly Gln
Val Ser 35 40 45 Pro Gly Tyr Ala Ser Pro Trp Ala Ala Pro Gly Ile
Pro Leu Lys Ala 50 55 60 Met Lys Trp Leu Leu Glu Lys His Ala Pro
Leu Ala Ile Lys Leu Thr 65 70 75 80 Ser Asp Pro Ser Gln Tyr Ala Trp
Met Leu Gln Met Leu Arg Asn Cys 85 90 95 Thr Ala Glu Arg Tyr Ala
Val Asn Lys Glu Arg Met Val Arg Leu Ser 100 105 110 Glu Tyr Ser Arg
Asp Cys Leu Asp Glu Leu Arg Ala Glu Thr Gly Ile 115 120 125 Ala Tyr
Glu Gly Arg Thr Leu Gly Thr Thr Gln Leu Phe Arg Thr Gln 130 135 140
Ala Gln Leu Asp Ala Ala Gly Lys Asp Ile Ala Val Leu Glu Arg Ser 145
150 155 160 Gly Val Pro Tyr Glu Val Leu Asp Arg Asp Gly Ile Ala Arg
Val Glu 165 170 175 Pro Ala Leu Ala Lys Val Ala Asp Lys Leu Val Gly
Ala Leu Arg Leu 180 185 190 Pro Asn Asp Gln Thr Gly Asp Cys Gln Leu
Phe Thr Thr Arg Leu Ala 195 200 205 Glu Met Ala Lys Gly Leu Gly Val
Glu Phe Arg Phe Gly Gln Asn Ile 210 215 220 Glu Arg Leu Asp Phe Ala
Gly Asp Arg Ile Asn Gly Val Leu Val Asn 225 230 235 240 Gly Glu Leu
Leu Thr Ala Asp His Tyr Val Leu Ala Leu Gly Ser Tyr 245 250 255 Ser
Pro Gln Leu Leu Lys Pro Leu Gly Ile Lys Ala Pro Val Tyr Pro 260 265
270 Leu Lys Gly Tyr Ser Leu Thr Val Pro Ile Thr Asn Pro Glu Met Ala
275 280 285 Pro Thr Ser Thr Ile Leu Asp Glu Thr Tyr Lys Val Ala Ile
Thr Arg 290 295 300 Phe Asp Gln Arg Ile Arg Val Gly Gly Met Ala Glu
Ile Ala Gly Phe 305 310 315 320 Asp Leu Ser Leu Asn Pro Arg Arg Arg
Glu Thr Leu Glu Met Ile Thr 325 330 335 Thr Asp Leu Tyr Pro Glu Gly
Gly Asp Ile Ser Gln Ala Thr Phe Trp 340 345 350 Thr Gly Leu Arg Pro
Ala Thr Pro Asp Gly Thr Pro Ile Val Gly Ala 355 360 365 Thr Arg Tyr
Arg Asn Leu Phe Leu Asn Thr Gly His Gly Thr Leu Gly 370 375 380 Trp
Thr Met Ala Cys Gly Ser Gly Arg Tyr Leu Ala Asp Leu Met Ala 385 390
395 400 Lys Lys Arg Pro Gln Ile Ser Thr Glu Gly Leu Asp Ile Ser Arg
Tyr 405 410 415 Ser Asn Ser Pro Glu Asn Ala Lys Asn Ala His Pro Ala
Pro Ala His 420 425 430 9 771 DNA Pseudomonas seruginosa 9
atggcactgg caaaacgcat catcccctgc ctcgacgtgg acaacggccg agtggtcaag
60 ggcgtcaagt tcgagaacat ccgcgacgcc ggcgacccgg tcgagatcgc
tcgccgctac 120 gacgagcagg gtgccgacga gatcaccttc ctcgatatca
ccgccagcgt cgacgggcgc 180 gacaccaccc tgcataccgt cgagcgcatg
gctagccagg tgttcattcc gctgaccgtg 240 ggcggcggcg tacgcagcgt
gcaggacatc cgcaacctgt tgaatgccgg cgcggacaag 300 gtctcgatca
acaccgccgc ggtgttcaac cccgagttcg tcggtgaggc cgccgaccgc 360
ttcggctcgc agtgcatcgt ggtcgccatc gacgcgaaga aggtttccgc cccgggcgag
420 gcgccgcgct gggaaatctt cacccatggc gggcgcaagc ccaccgggct
ggatgccgtg 480 ctctgggcga agaagatgga agacttgggc gctggcgaga
ttctcctgac cagcatggac 540 caggacggcg tgaagagcgg ttacgacctg
ggcgtgaccc gcgccatcag cgaggcggtg 600 aacgtgccgg tgatcgcttc
cggcggcgtc ggcaacctgg agcacctggc cgccggcatc 660 ctcgagggca
aggccgacgc ggtgctcgcg gcgagcatct tccacttcgg cgagtacacc 720
gtgccggaag ccaaggccta cctggccagc cgcggtatcg tggtgcgctg a 771 10 256
PRT Pseudomonas aeruginosa 10 Met Ala Leu Ala Lys Arg Ile Ile Pro
Cys Leu Asp Val Asp Asn Gly 1 5 10 15 Arg Val Val Lys Gly Val Lys
Phe Glu Asn Ile Arg Asp Ala Gly Asp 20 25 30 Pro Val Glu Ile Ala
Arg Arg Tyr Asp Glu Gln Gly Ala Asp Glu Ile 35 40 45 Thr Phe Leu
Asp Ile Thr Ala Ser Val Asp Gly Arg Asp Thr Thr Leu 50 55 60 His
Thr Val Glu Arg Met Ala Ser Gln Val Phe Ile Pro Leu Thr Val 65 70
75 80 Gly Gly Gly Val Arg Ser Val Gln Asp Ile Arg Asn Leu Leu Asn
Ala 85 90 95 Gly Ala Asp Lys Val Ser Ile Asn Thr Ala Ala Val Phe
Asn Pro Glu 100 105 110 Phe Val Gly Glu Ala Ala Asp Arg Phe Gly Ser
Gln Cys Ile Val Val 115 120 125 Ala Ile Asp Ala Lys Lys Val Ser Ala
Pro Gly Glu Ala Pro Arg Trp 130 135 140 Glu Ile Phe Thr His Gly Gly
Arg Lys Pro Thr Gly Leu Asp Ala Val 145 150 155 160 Leu Trp Ala Lys
Lys Met Glu Asp Leu Gly Ala Gly Glu Ile Leu Leu 165 170 175 Thr Ser
Met Asp Gln Asp Gly Val Lys Ser Gly Tyr Asp Leu Gly Val 180 185 190
Thr Arg Ala Ile Ser Glu Ala Val Asn Val Pro Val Ile Ala Ser Gly 195
200 205 Gly Val Gly Asn Leu Glu His Leu Ala Ala Gly Ile Leu Glu Gly
Lys 210 215 220 Ala Asp Ala Val Leu Ala Ala Ser Ile Phe His Phe Gly
Glu Tyr Thr 225 230
235 240 Val Pro Glu Ala Lys Ala Tyr Leu Ala Ser Arg Gly Ile Val Val
Arg 245 250 255 11 1035 DNA Pseudomonas aeruginosa 11 atgatcaagg
tcggcatcgt tggcggtacg ggttatacgg gcgtggaact gctgcgcctg 60
ctggcgcagc atccgcaggc ccgggtggaa gtgatcactt cgcgttccga ggcgggggtg
120 aaggtcgccg acatgtaccc gaacctgcga ggtcattatg acgacctgca
gttcagcgtg 180 ccggacgcgc agcgcctcgg cgcctgcgac gtggtgttct
tcgccacgcc gcacggcgtg 240 gcgcacgcgc tggctggcga actgctggac
gccgggaccc gggtcatcga tctgtccgct 300 gacttccgcc tggcggacgc
cgaggagtgg gcgcgctggt acggccagcc gcatggcgct 360 ccggcgctgc
tcgacgaggc tgtctacggc ctgccggaag tgaaccgcga gaagatccgc 420
caggcccgcc tgatcgccgt gccgggctgc tacccgaccg cgacccagct gggcctgatc
480 ccgctgctgg aagccggcct ggccgacgcc tcgcggctga tcgccgattg
caagtccggg 540 gtcagcggtg ccggtcgggg cgccaaggtt ggctcgctgt
tctgcgaggc gggcgaaagc 600 atgatggcct acgcggtcaa agggcatcgg
catctcccgg aaatcagcca gggcctgcgt 660 cgggcctccg gcggcgacgt
cgggctgacg ttcgtaccgc acctgacgcc aatgatccgc 720 ggtatccatg
caaccctcta tgcccatgtc gcggatcgct cggtcgacct ccaggcgttg 780
ttcgagaagc gctacgccga cgaacccttc gtcgacgtga tgccggccgg cagccatccg
840 gagacccgca gcgtgcgtgg cgccaatgtc tgccgaatcg ccgtgcatcg
cccccagggc 900 ggcgacctgg tggtggtgct gtcggtgatc gacaacctgg
tcaagggcgc ctcgggtcag 960 gcgctccaga acatgaacat cctgttcggg
ctggacgagc gcctgggcct ctcgcatgcg 1020 gccctgctcc cctga 1035 12 344
PRT Pseudomonas aeruginosa 12 Met Ile Lys Val Gly Ile Val Gly Gly
Thr Gly Tyr Thr Gly Val Glu 1 5 10 15 Leu Leu Arg Leu Leu Ala Gln
His Pro Gln Ala Arg Val Glu Val Ile 20 25 30 Thr Ser Arg Ser Glu
Ala Gly Val Lys Val Ala Asp Met Tyr Pro Asn 35 40 45 Leu Arg Gly
His Tyr Asp Asp Leu Gln Phe Ser Val Pro Asp Ala Gln 50 55 60 Arg
Leu Gly Ala Cys Asp Val Val Phe Phe Ala Thr Pro His Gly Val 65 70
75 80 Ala His Ala Leu Ala Gly Glu Leu Leu Asp Ala Gly Thr Arg Val
Ile 85 90 95 Asp Leu Ser Ala Asp Phe Arg Leu Ala Asp Ala Glu Glu
Trp Ala Arg 100 105 110 Trp Tyr Gly Gln Pro His Gly Ala Pro Ala Leu
Leu Asp Glu Ala Val 115 120 125 Tyr Gly Leu Pro Glu Val Asn Arg Glu
Lys Ile Arg Gln Ala Arg Leu 130 135 140 Ile Ala Val Pro Gly Cys Tyr
Pro Thr Ala Thr Gln Leu Gly Leu Ile 145 150 155 160 Pro Leu Leu Glu
Ala Gly Leu Ala Asp Ala Ser Arg Leu Ile Ala Asp 165 170 175 Cys Lys
Ser Gly Val Ser Gly Ala Gly Arg Gly Ala Lys Val Gly Ser 180 185 190
Leu Phe Cys Glu Ala Gly Glu Ser Met Met Ala Tyr Ala Val Lys Gly 195
200 205 His Arg His Leu Pro Glu Ile Ser Gln Gly Leu Arg Arg Ala Ser
Gly 210 215 220 Gly Asp Val Gly Leu Thr Phe Val Pro His Leu Thr Pro
Met Ile Arg 225 230 235 240 Gly Ile His Ala Thr Leu Tyr Ala His Val
Ala Asp Arg Ser Val Asp 245 250 255 Leu Gln Ala Leu Phe Glu Lys Arg
Tyr Ala Asp Glu Pro Phe Val Asp 260 265 270 Val Met Pro Ala Gly Ser
His Pro Glu Thr Arg Ser Val Arg Gly Ala 275 280 285 Asn Val Cys Arg
Ile Ala Val His Arg Pro Gln Gly Gly Asp Leu Val 290 295 300 Val Val
Leu Ser Val Ile Asp Asn Leu Val Lys Gly Ala Ser Gly Gln 305 310 315
320 Ala Leu Gln Asn Met Asn Ile Leu Phe Gly Leu Asp Glu Arg Leu Gly
325 330 335 Leu Ser His Ala Ala Leu Leu Pro 340 13 1644 DNA
Pseudomonas aeruginosa 13 gtgagcgaac tcattcgcgt acccgacatc
ggcaacggtg agggtgaagt catcgagctg 60 ctggtcaagc ccggcgacaa
ggtcgaggcc gatcagagcc tgctgaccct ggaatccgac 120 aaggccagca
tggaaatccc cagtcccaag gccggggtag tgaaaagcat caaggcgaag 180
gtcggcgaca ccttgaaaga aggtgacgaa atcctcgagc tggaagtgga aggcggcgaa
240 cagcctgccg aagccaaggc cgaggcagcg cccgcccaac cggaagcgcc
gaaagccgaa 300 gcgcctgctc ccgccccgag cgagagcaag ccggccgccc
ccgccgcggc cagcgtccag 360 gacatcaagg tcccggacat cggctcggcc
ggcaaggcca acgtcatcga agtgatggtc 420 aaggccggcg acacggtcga
ggccgaccag tcgctgatca ccctggaatc cgacaaggcc 480 agcatggaga
tcccctcgcc ggcctccggg gtggtggaaa gcgtctcgat caaggtcggt 540
gacgaagtcg gcaccggcga cctgatcctc aagctgaagg tggaaggcgc cgctccggca
600 gccgaagagc aaccggcagc cgctccggcc caggccgcgg cgcccgccgc
cgagcagaag 660 cccgccgcgg cggcccctgc gccagccaag gccgataccc
cggctccggt cggcgcaccc 720 agccgcgacg gcgccaaggt ccacgccggc
ccggcggtgc gcatgctggc gcgcgagttc 780 ggcgtcgagc tgagcgaagt
gaaagccagc ggtcccaagg gtcgcatcct caaggaagac 840 gtccaggtct
tcgtcaagga gcaactgcag cgcgccaagt ccggcggtgc cggcgccacc 900
ggcggagccg gcatcccgcc gatcccggaa gtcgacttca gcaagttcgg cgaagtggaa
960 gaagtggcga tgacccgcct gatgcaggtc ggcgccgcca acctgcatcg
cagctggctg 1020 aacgtgccgc acgtgaccca gttcgaccag tcggacatca
ccgacatgga agccttccgc 1080 gttgcccaga aggccgcggc ggagaaggcc
ggggtcaagc tgaccgtact gccgatcctg 1140 ctcaaggcct gcgcccacct
gctcaaggaa ctgccggact tcaacagttc gctggccccc 1200 agcggcaagg
cgctgatccg caagaagtac gtacacatcg gcttcgccgt ggacactccg 1260
gacggcctgc tggtcccggt gatccgcgat gtcgaccgga agagcctcct gcaactggcc
1320 gccgaggccg ccgacctggc cgacaaggcc cgcaacaaga agctctcggc
cgatgccatg 1380 cagggcgcct gcttcaccat ctccagtctc ggccacatcg
gcggcaccgg cttcacgccg 1440 atcgtcaacg cgccggaagt ggcgatcctc
ggtgtgtcca aggcgaccat gcagccggta 1500 tgggacggca aggccttcca
gccgcgcctg atgctgccgc tgtcgctgtc ctacgaccat 1560 cgcgtgatca
acggtgccgc cgcggcgcgc ttcaccaagc gcctgggcga gctgctggcg 1620
gacatccgca ccctgctcct gtaa 1644 14 547 PRT Pseudomonas aeruginosa
14 Met Ser Glu Leu Ile Arg Val Pro Asp Ile Gly Asn Gly Glu Gly Glu
1 5 10 15 Val Ile Glu Leu Leu Val Lys Pro Gly Asp Lys Val Glu Ala
Asp Gln 20 25 30 Ser Leu Leu Thr Leu Glu Ser Asp Lys Ala Ser Met
Glu Ile Pro Ser 35 40 45 Pro Lys Ala Gly Val Val Lys Ser Ile Lys
Ala Lys Val Gly Asp Thr 50 55 60 Leu Lys Glu Gly Asp Glu Ile Leu
Glu Leu Glu Val Glu Gly Gly Glu 65 70 75 80 Gln Pro Ala Glu Ala Lys
Ala Glu Ala Ala Pro Ala Gln Pro Glu Ala 85 90 95 Pro Lys Ala Glu
Ala Pro Ala Pro Ala Pro Ser Glu Ser Lys Pro Ala 100 105 110 Ala Pro
Ala Ala Ala Ser Val Gln Asp Ile Lys Val Pro Asp Ile Gly 115 120 125
Ser Ala Gly Lys Ala Asn Val Ile Glu Val Met Val Lys Ala Gly Asp 130
135 140 Thr Val Glu Ala Asp Gln Ser Leu Ile Thr Leu Glu Ser Asp Lys
Ala 145 150 155 160 Ser Met Glu Ile Pro Ser Pro Ala Ser Gly Val Val
Glu Ser Val Ser 165 170 175 Ile Lys Val Gly Asp Glu Val Gly Thr Gly
Asp Leu Ile Leu Lys Leu 180 185 190 Lys Val Glu Gly Ala Ala Pro Ala
Ala Glu Glu Gln Pro Ala Ala Ala 195 200 205 Pro Ala Gln Ala Ala Ala
Pro Ala Ala Glu Gln Lys Pro Ala Ala Ala 210 215 220 Ala Pro Ala Pro
Ala Lys Ala Asp Thr Pro Ala Pro Val Gly Ala Pro 225 230 235 240 Ser
Arg Asp Gly Ala Lys Val His Ala Gly Pro Ala Val Arg Met Leu 245 250
255 Ala Arg Glu Phe Gly Val Glu Leu Ser Glu Val Lys Ala Ser Gly Pro
260 265 270 Lys Gly Arg Ile Leu Lys Glu Asp Val Gln Val Phe Val Lys
Glu Gln 275 280 285 Leu Gln Arg Ala Lys Ser Gly Gly Ala Gly Ala Thr
Gly Gly Ala Gly 290 295 300 Ile Pro Pro Ile Pro Glu Val Asp Phe Ser
Lys Phe Gly Glu Val Glu 305 310 315 320 Glu Val Ala Met Thr Arg Leu
Met Gln Val Gly Ala Ala Asn Leu His 325 330 335 Arg Ser Trp Leu Asn
Val Pro His Val Thr Gln Phe Asp Gln Ser Asp 340 345 350 Ile Thr Asp
Met Glu Ala Phe Arg Val Ala Gln Lys Ala Ala Ala Glu 355 360 365 Lys
Ala Gly Val Lys Leu Thr Val Leu Pro Ile Leu Leu Lys Ala Cys 370 375
380 Ala His Leu Leu Lys Glu Leu Pro Asp Phe Asn Ser Ser Leu Ala Pro
385 390 395 400 Ser Gly Lys Ala Leu Ile Arg Lys Lys Tyr Val His Ile
Gly Phe Ala 405 410 415 Val Asp Thr Pro Asp Gly Leu Leu Val Pro Val
Ile Arg Asp Val Asp 420 425 430 Arg Lys Ser Leu Leu Gln Leu Ala Ala
Glu Ala Ala Asp Leu Ala Asp 435 440 445 Lys Ala Arg Asn Lys Lys Leu
Ser Ala Asp Ala Met Gln Gly Ala Cys 450 455 460 Phe Thr Ile Ser Ser
Leu Gly His Ile Gly Gly Thr Gly Phe Thr Pro 465 470 475 480 Ile Val
Asn Ala Pro Glu Val Ala Ile Leu Gly Val Ser Lys Ala Thr 485 490 495
Met Gln Pro Val Trp Asp Gly Lys Ala Phe Gln Pro Arg Leu Met Leu 500
505 510 Pro Leu Ser Leu Ser Tyr Asp His Arg Val Ile Asn Gly Ala Ala
Ala 515 520 525 Ala Arg Phe Thr Lys Arg Leu Gly Glu Leu Leu Ala Asp
Ile Arg Thr 530 535 540 Leu Leu Leu 545 15 996 DNA Pseudomonas
aeruginosa 15 atgagttggc tgactcccgc tctggtcacc atcatcctca
ccgtggtcaa ggccatcgtg 60 gtgctgctcg ccgtggtcat ctgcggcgcc
ctgctaagct gggtcgagcg ccgcctgctc 120 ggcctctggc aggaccgcta
cggccccaac cgggtcggtc cgttcggtgc gttccagctc 180 ggcgcggaca
tggtcaagat gttcttcaag gaggactgga ccccgccgtt cgccgacaag 240
atgatcttca ccctggcccc ggtaatcgcg atgggcgccc tgctcgtcgc cttcgccatc
300 gtgccgatca cccccacctg gggcgtggcg gacctgaaca tcggcatcct
gttcttcttc 360 gccatggccg gcctgacggt gtacgccgtg ctgttcgccg
gctggtcgag caacaacaag 420 ttcgccctgc tcggcagcct gcgcgcctcg
gcccagacca tctcctacga ggtgttcctg 480 gccctgtcgc tgatgggcat
cgtcgcccag gtcggctcgt tcaacatgcg cgacatcgtc 540 cagtaccaga
tcgacaacgt ctggttcatc attccgcagt tcttcggctt ctgcaccttc 600
atcatcgccg gcgtcgccgt gacccaccgt cacccgttcg accagccgga agcggagcag
660 gaactggcgg acggctacca catcgagtac gccgggatga aatggggcat
gttcttcgtc 720 ggcgagtaca tcggcatcgt actggtctcg gcgctgctgg
cgaccctgtt cttcggcggc 780 tggcacggtc cgttcctgga caccctgccc
tggctgtcgt tcttctactt cgccgccaag 840 accggcttct tcatcatgct
cttcatcctg atccgcgcct cgctgccgcg tccgcgctat 900 gaccaggtga
tggcgttcag ctggaaggtg tgcctgccgc tgaccctgat caacctgctg 960
gtgaccggcg cgctcgtgct ggccgcggcc cagtaa 996 16 331 PRT Pseudomonas
aeruginosa 16 Met Ser Trp Leu Thr Pro Ala Leu Val Thr Ile Ile Leu
Thr Val Val 1 5 10 15 Lys Ala Ile Val Val Leu Leu Ala Val Val Ile
Cys Gly Ala Leu Leu 20 25 30 Ser Trp Val Glu Arg Arg Leu Leu Gly
Leu Trp Gln Asp Arg Tyr Gly 35 40 45 Pro Asn Arg Val Gly Pro Phe
Gly Ala Phe Gln Leu Gly Ala Asp Met 50 55 60 Val Lys Met Phe Phe
Lys Glu Asp Trp Thr Pro Pro Phe Ala Asp Lys 65 70 75 80 Met Ile Phe
Thr Leu Ala Pro Val Ile Ala Met Gly Ala Leu Leu Val 85 90 95 Ala
Phe Ala Ile Val Pro Ile Thr Pro Thr Trp Gly Val Ala Asp Leu 100 105
110 Asn Ile Gly Ile Leu Phe Phe Phe Ala Met Ala Gly Leu Thr Val Tyr
115 120 125 Ala Val Leu Phe Ala Gly Trp Ser Ser Asn Asn Lys Phe Ala
Leu Leu 130 135 140 Gly Ser Leu Arg Ala Ser Ala Gln Thr Ile Ser Tyr
Glu Val Phe Leu 145 150 155 160 Ala Leu Ser Leu Met Gly Ile Val Ala
Gln Val Gly Ser Phe Asn Met 165 170 175 Arg Asp Ile Val Gln Tyr Gln
Ile Asp Asn Val Trp Phe Ile Ile Pro 180 185 190 Gln Phe Phe Gly Phe
Cys Thr Phe Ile Ile Ala Gly Val Ala Val Thr 195 200 205 His Arg His
Pro Phe Asp Gln Pro Glu Ala Glu Gln Glu Leu Ala Asp 210 215 220 Gly
Tyr His Ile Glu Tyr Ala Gly Met Lys Trp Gly Met Phe Phe Val 225 230
235 240 Gly Glu Tyr Ile Gly Ile Val Leu Val Ser Ala Leu Leu Ala Thr
Leu 245 250 255 Phe Phe Gly Gly Trp His Gly Pro Phe Leu Asp Thr Leu
Pro Trp Leu 260 265 270 Ser Phe Phe Tyr Phe Ala Ala Lys Thr Gly Phe
Phe Ile Met Leu Phe 275 280 285 Ile Leu Ile Arg Ala Ser Leu Pro Arg
Pro Arg Tyr Asp Gln Val Met 290 295 300 Ala Phe Ser Trp Lys Val Cys
Leu Pro Leu Thr Leu Ile Asn Leu Leu 305 310 315 320 Val Thr Gly Ala
Leu Val Leu Ala Ala Ala Gln 325 330 17 7347 DNA Pseudomonas
aeruginosa 17 gtgcaagcac tcatagagaa ggtgggctcc ctttcccccc
aggaaaggaa ggcattggct 60 gtcctgctca agcagcaagg tgtcaatctc
ttcgagatcg cgccagtgtt caagcgccag 120 gacggcgagc ccctgcggct
ctcctatgcc caggagcgac agtggtttct ctggcaactg 180 gagccggaaa
gcgcggccta ccatatcccg agtgtcttgc gtctacgtgg gcggctggac 240
ctggatgccc tgcaacgcag cttcgacagc ctggttgcgc ggcacgagac cctacgcacc
300 cgttttcgcc tcgacggcga cgaggcgcgc caggagatcg ccgcatccat
ggcattgccg 360 ttggatatcg tcgcgttggg gccgctcgag gagggcgccc
tcgctcggca ggtcgagacg 420 acgatcgcgc ggccgttcga cctggagcgt
gggccgctgc tgcgggtgag cctgttgcgg 480 ctggccgagg acgaccatgt
gctggtgctg gtccagcatc acatcgtgtc cgacggttgg 540 tcgatgcagg
tgatggtcga ggaactggtc cagctctatg ccgcctatag tcgagggctc 600
gaggtagcgc tgccggcttt gccgatccag tacgcggact acgccctgtg gcagcgcagc
660 tggatggagg ccggggaaaa ggagcgccag ttggcgtact ggaccggcct
gctgggcggc 720 gagcagccgg tgctggagtt gccgttcgac cggccgcgcc
cggttcggca aagccatcgt 780 ggtgcccagt tcatcctgga actggatatt
gatctgtccc aggcgctcag gcgcgtggcc 840 cagcaggagg gggctactgc
cttcgccctg ttgctggctt cgttccaggc gctgctgtat 900 cgctacagcg
ggcaggcgga tatccgtgtc ggcgtgccga tcgccaatcg caaccgcgtg 960
gagaccgagc ggctgatcgg cttcttcgtc aacacccagg tgctcaaggc cgacctggac
1020 ggtcggatgg gcttcgacga gctgctggcc caggcccgcc aacgcgcgct
ggaggcccag 1080 gcgcaccagg acctgccgtt cgagcaactg gtggaggcct
tgcagccgga gcgcagtctt 1140 agccacaacc cgctgttcca ggtgctgttc
aactaccaga gcgaagcccg tggcaacggc 1200 caggcattcc gcttcgacga
gttacagatg gaaagcgtgc agttcgacag ccggacggcg 1260 cagttcgact
tgacgttgga cctgacggac gaagagcagc gtttttgcgc cgttttcgac 1320
tacgccaccg acctgttcga cgcctccacc gtggaacgcc tggccggcca ttggcgcaac
1380 ctgttgcgcg gcatcgtcgc caacccacga cagcggctcg gcgagttgcc
gctgctggat 1440 gcgccggagc gccggcagac cctctccgaa tggaacccgg
cccagcgcga gtgcgcggtg 1500 cagggcacct tgcagcagcg tttcgaggaa
caggcgcggc aacggccaca ggcggttgcg 1560 ctgatcctcg acgaacaacg
gttgagctac ggcgaactga atgcgcgggc caatcgcctg 1620 gcgcactgcc
tgatcgcccg tggcgttggc gcggacgtgc cggtcgggct ggcgctggag 1680
cgttcgctgg acatgctggt cggcttgctg gcgatcctca aggccggcgg cgcctacctg
1740 ccgttggacc cggcggcgcc agaggagcgc ctggcgcata tcctcgacga
cagtggggta 1800 cggctgctgc tgacccaggg gcatctgctc gagcgcctgc
cacggcaggc gggggtggag 1860 gtgctggcca tcgacggact ggtgctggac
ggctacgccg agagcgatcc gctcccgacg 1920 ctatcggcgg acaacctggc
ctacgtgatc tatacctcgg gctcgaccgg caagcccaag 1980 ggcacattgc
tcacccaccg caacgcgctg cgcctgttca gcgccaccga ggcctggttc 2040
ggcttcgacg agcgggacgt gtggacattg ttccattcct acgccttcga tttctcggtc
2100 tgggaaatct tcggcgcgct gctctatggc gggtgcctgg tgattgtgcc
gcaatgggtg 2160 agccgttcgc cggaagactt ctaccgtctg ctgtgccgcg
aaggcgtgac ggtgctcaac 2220 cagacgccgt cggcgttcaa gcaactgatg
gcggtggcct gttccgccga catggcgacg 2280 cagcagccgg cgctgcgcta
cgtgatcttc ggtggcgagg cgctggatct gcagagcctg 2340 cggccgtggt
tccagcgctt cggcgatcgc cagccgcaac tggtgaacat gtacggcatc 2400
accgagacca cggtgcacgt aacctaccgt ccggtgagcg aggccgacct ggaaggtggc
2460 ctggtcagtc cgattggcgg gaccatcccg gacctgtcct ggtacatcct
cgaccgtgac 2520 ctgaacccgg tgccgcgcgg cgcggtgggc gagctgtaca
tcggtcgcgc cgggctggcg 2580 cgcggctacc tgaggcggcc cgggttgagt
gccacccgct tcgtgccgaa cccgttcccc 2640 ggcggcgccg gcgagcggct
gtaccgtacc ggcgacctgg cacggttcca ggcggatggc 2700 aatatcgagt
acatcgggcg tatcgaccac caggtgaagg ttcgcggctt ccgtatcgaa 2760
ctgggcgaga tcgaagcggc gctcgccggt ctggccgggg tacgcgatgc cgtggtgctg
2820 gcccatgacg gagtcggcgg cacgcaactg gtgggatacg tggtggcgga
ctcggcggag 2880 gatgccgagc gtctgcggga gtcgctgcgg gagtcgctga
agcggcacct gccggactac 2940 atggtgccgg cgcacctgat gctgctggag
cggatgccgc tgacggtcaa tggcaagctc 3000 gaccggcagg cgttgccgca
accggatgcg agcctgtcgc aacaggccta tcgagcgccc 3060 ggtagcgagc
tggagcagcg catcgcagcg atctggtcgg agatcctggg agtggaacgg 3120
gtcggcctgg acgacaactt cttcgaactg ggcggtcatt cgttgctggc tacccgggtg
3180 atttctcggg ttcgccagga gcagcagttg gacgcaagcc tgaaggcgtt
gttcgagcgg 3240 ccggttctgg aagcgttcgc ccagggattg gaacgcacga
cggatgcggt ctcgacgata 3300 ccgcttgccg atcggcagca accgttggca
ctgtccttcg ctcaggagcg tcagtggttc 3360 ctctggcaac tggagccgga
aagcgcggcc taccatattc cgagtgcctt gcgcctacgc 3420
gggcggctgg acgtggatgc cttgcaacgc agcttcgaca gcctggtcgc gcggcatgaa
3480 accttgcgta cccgcttccg gctggaggga gggcgttcgt accagcaggt
acaacctgcg 3540 gttagcgttt ccatcgagcg ggaacagttc ggtgaagaag
gcctgatcga acggatacag 3600 gccatcgttg tgcagccatt cgacctggaa
cgggggccgc tgctgcgggt gaacctgttg 3660 caactggccg aggacgacca
tgtactggtg ctggtccagc accacatcgt gtccgatggt 3720 tggtcgatgc
aggtgatggt cgaggaactg gtccagctct atgccgccta tagccaaggg 3780
ctcgacgtgg tgttgccagc cctgccgatc cagtacgcgg actacgccct gtggcagcgc
3840 agctggatgg aggcggggga aaaggagcgc cagttggcgt actggaccgg
cctgctgggc 3900 ggcgagcagc cggtgctgga gttgccgttc gatcggccgc
gtccggcccg gcagagccat 3960 cgtggcgcgc agttgggttt cgagctatcg
cgggaactgg tcgaggccgt gagagccttg 4020 gcccagcgtg aaggcgccag
tagtttcatg ctgttgctgg cctcgttcca ggcgctgttg 4080 tatcgctaca
gcgggcaggc ggatatccgt gtcggtgtgc cgatcgccaa tcgcaaccgc 4140
gtggagaccg agcggctgat cggcttcttc gtcaacaccc aggtgctcaa ggccgacctg
4200 gacggtcgga tgggcttcga cgagctgctg gcccaggccc gccaacgcgc
gctggaggcc 4260 caggcgcacc aggacctgcc gttcgagcaa ctggtggaag
ccttgcagcc ggagcgcaat 4320 gccagccaca acccactgtt ccaggtgctg
ttcaaccatc agagcgagat acgctcggtg 4380 acgcccgagg ttcagttgga
ggacctgcgt ctggaaggcc tggcctggga cggccagact 4440 gcgcagttcg
acctgacgct ggatattcag gaagacgaaa acggcatctg ggcctccttc 4500
gactatgcca ccgatctgtt cgacgcctcc accgtggaac gcctggccgg ccattggcgc
4560 aacctgttgc gcggcatcgt cgccaaccca cgacagcggc tcggcgagtt
gccgctgctg 4620 gatgcgccgg agcgccggca gaccctctcc gaatggaacc
cggcccagcg cgagtgcgcg 4680 gtgcagggca ccttgcagca gcgtttcgag
gagcaggcgc ggcaacggcc acaggcggtt 4740 gcgctgatcc tcgacgaaca
acggttgagc tacggcgaac tgaatgcgcg ggccaatcgc 4800 ctggcgcact
gcctgatcgc tcgcggcgtt ggcgcggacg tgccggtcgg gctggcgctg 4860
gagcgttcgc tggacatgct ggtcggcttg ctggcgatcc tcaaggccgg cggcgcctac
4920 ctgccgttgg acccggcggc gccagaggag cgcctggcgc atatcctcga
cgacagtggg 4980 gtacggctgc tgctgaccca ggggcatctg ctcgagcgcc
tgccgcggca ggcgggggtg 5040 gaggtgctgg ccatcgacgg actggtgctg
gacggctacg ccgagagcga tccgctcccg 5100 acgctatcgg cggacaacct
ggcctacgtg atctatacct cgggctcgac cggcaagccc 5160 aagggcacgt
tgctcaccca ccgcaacgcg ctgcgcctgt tcagcgccac cgaggcctgg 5220
ttcggcttcg acgagcggga cgtgtggacg ttgttccatt cctacgcctt cgatttctcg
5280 gtctgggaaa tcttcggcgc gctgctctat ggcgggcgcc tggtgatcgt
gccgcaatgg 5340 gtgagccgtt cgccggaaga cttctaccgt ctgctgtgcc
gcgaaggcgt gacggtgctc 5400 aaccagacgc cgtcggcgtt caagcaactg
atggcggtgg cctgttccgc cgacatggcg 5460 acgcagcagc cggcgctgcg
ctacgtgatc ttcggtggcg aggcgctgga tctgcagagc 5520 ctgcggccgt
ggttccagcg ctttggcgat cgccagccgc aactggtgaa catgtacggc 5580
atcaccgaga ccacggtaca cgtaacctac cgtccggtga gcgaagccga cctgaagggt
5640 ggcctggtca gtccgatcgg cgggaccatc ccggacctgt cctggtacat
cctcgaccgt 5700 gacctgaacc cggtgccgcg cggcgcggtg ggcgagctgt
acatcggtcg cgccggtctg 5760 gcgcgcggct acctgaggcg gcccgggttg
agtgccaccc gcttcgtgcc gaacccgttc 5820 cccggcggtg ccggcgagcg
gctgtaccgt accggcgacc tggcacggtt ccaggcggat 5880 ggcaatatcg
agtacatcgg gcgtatcgac caccaggtga aggttcgcgg cttccgtatc 5940
gaactgggtg agatcgaagc ggcgctcgcc ggtctggccg gggtacgcga tgccgtggtg
6000 ctggcccatg acggggtcgg cggcacgcaa ctggtgggat acgtggtggc
ggactcggcg 6060 gaggatgccg agcgtctgcg ggagtcgctg cgggagtcgc
tgaagcggca cctgccggac 6120 tacatggtgc cggcgcacct gatgctgctg
gagcggatgc cgctgacggt caatggcaag 6180 ctcgaccggc aggcgttgcc
gcaaccggat gcgagcttgt cgcagcaggc ctatcgagcg 6240 cccggtagcg
agctggagca gcgcatcgca gcgatctggg cggagatcct gggagtggaa 6300
cgggtcggcc tggacgacaa cttcttcgaa ctgggcggtc actcattgtt gctgctgatg
6360 ctcaaggagc ggatcggcga tacctgccag gctacgctga gcatcagcca
actgatgacc 6420 catgccagcg tcgcggaaca ggcggcatgc atcgaggggc
aggcgcgtga gtcgttgctg 6480 gtgccgctca acggcaggcg cgaaggttcg
ccgctgttca tgttccatcc gagtttcggc 6540 tctgtgcact gttacaagac
cctcgccatg gcgctgcggg atcgtcatcc ggtcaagggt 6600 gttgtctgcc
gtgccctgct gggcgctggt cgcgaggtgc cggagtggga cgatatggtt 6660
gcggaatacg ccgagcaatt gctgcaggag caccccgaag gggttttcaa cctggcggga
6720 tggtcgctcg gcggcaacct ggcgatggat gtcgcggccc ggctggagca
gcgtgggcgg 6780 caggtggctt tcgtcggctg gatcgatgca ccggcaccgg
tcagggtcga agcgttctgg 6840 aacgagatcg ggccgacgcc ggaggcagtc
ccgaacctat ccgtgggcga gatgcgggtg 6900 gaactgctcg gtgtcatgtt
tccggagcgg gccgagcata tcgaacgggc ctggtcatcg 6960 atctgctccg
ccacgacgga cgatgagcag cgctggacga ggatgagcga ctgggcggaa 7020
gcggagatcg gcgccgagtt cgcgacactg cgcagcgaaa tcgcacagag caacgaactg
7080 gaagtgtcct gggagttgaa acagatcctc gacgagcgcc tgaaagcgat
ggattacccg 7140 cgtctgacgg cgaaggtcag cctctggtgg gccgcgcgca
gcaccaatgc catccagcgg 7200 agcgcggtgg agcgctcgat ggccgaggcg
atcggggctg agcgtgtcga accggtgcgg 7260 gtgctggata cccggcacga
caagatcatc gaccaccctg agtttgtgca gagcttccgg 7320 gccgccctgg
agcgtgccgg gcgctga 7347 18 2448 PRT Pseudomonas aeruginosa 18 Met
Gln Ala Leu Ile Glu Lys Val Gly Ser Leu Ser Pro Gln Glu Arg 1 5 10
15 Lys Ala Leu Ala Val Leu Leu Lys Gln Gln Gly Val Asn Leu Phe Glu
20 25 30 Ile Ala Pro Val Phe Lys Arg Gln Asp Gly Glu Pro Leu Arg
Leu Ser 35 40 45 Tyr Ala Gln Glu Arg Gln Trp Phe Leu Trp Gln Leu
Glu Pro Glu Ser 50 55 60 Ala Ala Tyr His Ile Pro Ser Val Leu Arg
Leu Arg Gly Arg Leu Asp 65 70 75 80 Leu Asp Ala Leu Gln Arg Ser Phe
Asp Ser Leu Val Ala Arg His Glu 85 90 95 Thr Leu Arg Thr Arg Phe
Arg Leu Asp Gly Asp Glu Ala Arg Gln Glu 100 105 110 Ile Ala Ala Ser
Met Ala Leu Pro Leu Asp Ile Val Ala Leu Gly Pro 115 120 125 Leu Glu
Glu Gly Ala Leu Ala Arg Gln Val Glu Thr Thr Ile Ala Arg 130 135 140
Pro Phe Asp Leu Glu Arg Gly Pro Leu Leu Arg Val Ser Leu Leu Arg 145
150 155 160 Leu Ala Glu Asp Asp His Val Leu Val Leu Val Gln His His
Ile Val 165 170 175 Ser Asp Gly Trp Ser Met Gln Val Met Val Glu Glu
Leu Val Gln Leu 180 185 190 Tyr Ala Ala Tyr Ser Arg Gly Leu Glu Val
Ala Leu Pro Ala Leu Pro 195 200 205 Ile Gln Tyr Ala Asp Tyr Ala Leu
Trp Gln Arg Ser Trp Met Glu Ala 210 215 220 Gly Glu Lys Glu Arg Gln
Leu Ala Tyr Trp Thr Gly Leu Leu Gly Gly 225 230 235 240 Glu Gln Pro
Val Leu Glu Leu Pro Phe Asp Arg Pro Arg Pro Val Arg 245 250 255 Gln
Ser His Arg Gly Ala Gln Phe Ile Leu Glu Leu Asp Ile Asp Leu 260 265
270 Ser Gln Ala Leu Arg Arg Val Ala Gln Gln Glu Gly Ala Thr Ala Phe
275 280 285 Ala Leu Leu Leu Ala Ser Phe Gln Ala Leu Leu Tyr Arg Tyr
Ser Gly 290 295 300 Gln Ala Asp Ile Arg Val Gly Val Pro Ile Ala Asn
Arg Asn Arg Val 305 310 315 320 Glu Thr Glu Arg Leu Ile Gly Phe Phe
Val Asn Thr Gln Val Leu Lys 325 330 335 Ala Asp Leu Asp Gly Arg Met
Gly Phe Asp Glu Leu Leu Ala Gln Ala 340 345 350 Arg Gln Arg Ala Leu
Glu Ala Gln Ala His Gln Asp Leu Pro Phe Glu 355 360 365 Gln Leu Val
Glu Ala Leu Gln Pro Glu Arg Ser Leu Ser His Asn Pro 370 375 380 Leu
Phe Gln Val Leu Phe Asn Tyr Gln Ser Glu Ala Arg Gly Asn Gly 385 390
395 400 Gln Ala Phe Arg Phe Asp Glu Leu Gln Met Glu Ser Val Gln Phe
Asp 405 410 415 Ser Arg Thr Ala Gln Phe Asp Leu Thr Leu Asp Leu Thr
Asp Glu Glu 420 425 430 Gln Arg Phe Cys Ala Val Phe Asp Tyr Ala Thr
Asp Leu Phe Asp Ala 435 440 445 Ser Thr Val Glu Arg Leu Ala Gly His
Trp Arg Asn Leu Leu Arg Gly 450 455 460 Ile Val Ala Asn Pro Arg Gln
Arg Leu Gly Glu Leu Pro Leu Leu Asp 465 470 475 480 Ala Pro Glu Arg
Arg Gln Thr Leu Ser Glu Trp Asn Pro Ala Gln Arg 485 490 495 Glu Cys
Ala Val Gln Gly Thr Leu Gln Gln Arg Phe Glu Glu Gln Ala 500 505 510
Arg Gln Arg Pro Gln Ala Val Ala Leu Ile Leu Asp Glu Gln Arg Leu 515
520 525 Ser Tyr Gly Glu Leu Asn Ala Arg Ala Asn Arg Leu Ala His Cys
Leu 530 535 540 Ile Ala Arg Gly Val Gly Ala Asp Val Pro Val Gly Leu
Ala Leu Glu 545 550 555 560 Arg Ser Leu Asp Met Leu Val Gly Leu Leu
Ala Ile Leu Lys Ala Gly 565 570 575 Gly Ala Tyr Leu Pro Leu Asp Pro
Ala Ala Pro Glu Glu Arg Leu Ala 580 585 590 His Ile Leu Asp Asp Ser
Gly Val Arg Leu Leu Leu Thr Gln Gly His 595 600 605 Leu Leu Glu Arg
Leu Pro Arg Gln Ala Gly Val Glu Val Leu Ala Ile 610 615 620 Asp Gly
Leu Val Leu Asp Gly Tyr Ala Glu Ser Asp Pro Leu Pro Thr 625 630 635
640 Leu Ser Ala Asp Asn Leu Ala Tyr Val Ile Tyr Thr Ser Gly Ser Thr
645 650 655 Gly Lys Pro Lys Gly Thr Leu Leu Thr His Arg Asn Ala Leu
Arg Leu 660 665 670 Phe Ser Ala Thr Glu Ala Trp Phe Gly Phe Asp Glu
Arg Asp Val Trp 675 680 685 Thr Leu Phe His Ser Tyr Ala Phe Asp Phe
Ser Val Trp Glu Ile Phe 690 695 700 Gly Ala Leu Leu Tyr Gly Gly Cys
Leu Val Ile Val Pro Gln Trp Val 705 710 715 720 Ser Arg Ser Pro Glu
Asp Phe Tyr Arg Leu Leu Cys Arg Glu Gly Val 725 730 735 Thr Val Leu
Asn Gln Thr Pro Ser Ala Phe Lys Gln Leu Met Ala Val 740 745 750 Ala
Cys Ser Ala Asp Met Ala Thr Gln Gln Pro Ala Leu Arg Tyr Val 755 760
765 Ile Phe Gly Gly Glu Ala Leu Asp Leu Gln Ser Leu Arg Pro Trp Phe
770 775 780 Gln Arg Phe Gly Asp Arg Gln Pro Gln Leu Val Asn Met Tyr
Gly Ile 785 790 795 800 Thr Glu Thr Thr Val His Val Thr Tyr Arg Pro
Val Ser Glu Ala Asp 805 810 815 Leu Glu Gly Gly Leu Val Ser Pro Ile
Gly Gly Thr Ile Pro Asp Leu 820 825 830 Ser Trp Tyr Ile Leu Asp Arg
Asp Leu Asn Pro Val Pro Arg Gly Ala 835 840 845 Val Gly Glu Leu Tyr
Ile Gly Arg Ala Gly Leu Ala Arg Gly Tyr Leu 850 855 860 Arg Arg Pro
Gly Leu Ser Ala Thr Arg Phe Val Pro Asn Pro Phe Pro 865 870 875 880
Gly Gly Ala Gly Glu Arg Leu Tyr Arg Thr Gly Asp Leu Ala Arg Phe 885
890 895 Gln Ala Asp Gly Asn Ile Glu Tyr Ile Gly Arg Ile Asp His Gln
Val 900 905 910 Lys Val Arg Gly Phe Arg Ile Glu Leu Gly Glu Ile Glu
Ala Ala Leu 915 920 925 Ala Gly Leu Ala Gly Val Arg Asp Ala Val Val
Leu Ala His Asp Gly 930 935 940 Val Gly Gly Thr Gln Leu Val Gly Tyr
Val Val Ala Asp Ser Ala Glu 945 950 955 960 Asp Ala Glu Arg Leu Arg
Glu Ser Leu Arg Glu Ser Leu Lys Arg His 965 970 975 Leu Pro Asp Tyr
Met Val Pro Ala His Leu Met Leu Leu Glu Arg Met 980 985 990 Pro Leu
Thr Val Asn Gly Lys Leu Asp Arg Gln Ala Leu Pro Gln Pro 995 1000
1005 Asp Ala Ser Leu Ser Gln Gln Ala Tyr Arg Ala Pro Gly Ser Glu
1010 1015 1020 Leu Glu Gln Arg Ile Ala Ala Ile Trp Ser Glu Ile Leu
Gly Val 1025 1030 1035 Glu Arg Val Gly Leu Asp Asp Asn Phe Phe Glu
Leu Gly Gly His 1040 1045 1050 Ser Leu Leu Ala Thr Arg Val Ile Ser
Arg Val Arg Gln Glu Gln 1055 1060 1065 Gln Leu Asp Ala Ser Leu Lys
Ala Leu Phe Glu Arg Pro Val Leu 1070 1075 1080 Glu Ala Phe Ala Gln
Gly Leu Glu Arg Thr Thr Asp Ala Val Ser 1085 1090 1095 Thr Ile Pro
Leu Ala Asp Arg Gln Gln Pro Leu Ala Leu Ser Phe 1100 1105 1110 Ala
Gln Glu Arg Gln Trp Phe Leu Trp Gln Leu Glu Pro Glu Ser 1115 1120
1125 Ala Ala Tyr His Ile Pro Ser Ala Leu Arg Leu Arg Gly Arg Leu
1130 1135 1140 Asp Val Asp Ala Leu Gln Arg Ser Phe Asp Ser Leu Val
Ala Arg 1145 1150 1155 His Glu Thr Leu Arg Thr Arg Phe Arg Leu Glu
Gly Gly Arg Ser 1160 1165 1170 Tyr Gln Gln Val Gln Pro Ala Val Ser
Val Ser Ile Glu Arg Glu 1175 1180 1185 Gln Phe Gly Glu Glu Gly Leu
Ile Glu Arg Ile Gln Ala Ile Val 1190 1195 1200 Val Gln Pro Phe Asp
Leu Glu Arg Gly Pro Leu Leu Arg Val Asn 1205 1210 1215 Leu Leu Gln
Leu Ala Glu Asp Asp His Val Leu Val Leu Val Gln 1220 1225 1230 His
His Ile Val Ser Asp Gly Trp Ser Met Gln Val Met Val Glu 1235 1240
1245 Glu Leu Val Gln Leu Tyr Ala Ala Tyr Ser Gln Gly Leu Asp Val
1250 1255 1260 Val Leu Pro Ala Leu Pro Ile Gln Tyr Ala Asp Tyr Ala
Leu Trp 1265 1270 1275 Gln Arg Ser Trp Met Glu Ala Gly Glu Lys Glu
Arg Gln Leu Ala 1280 1285 1290 Tyr Trp Thr Gly Leu Leu Gly Gly Glu
Gln Pro Val Leu Glu Leu 1295 1300 1305 Pro Phe Asp Arg Pro Arg Pro
Ala Arg Gln Ser His Arg Gly Ala 1310 1315 1320 Gln Leu Gly Phe Glu
Leu Ser Arg Glu Leu Val Glu Ala Val Arg 1325 1330 1335 Ala Leu Ala
Gln Arg Glu Gly Ala Ser Ser Phe Met Leu Leu Leu 1340 1345 1350 Ala
Ser Phe Gln Ala Leu Leu Tyr Arg Tyr Ser Gly Gln Ala Asp 1355 1360
1365 Ile Arg Val Gly Val Pro Ile Ala Asn Arg Asn Arg Val Glu Thr
1370 1375 1380 Glu Arg Leu Ile Gly Phe Phe Val Asn Thr Gln Val Leu
Lys Ala 1385 1390 1395 Asp Leu Asp Gly Arg Met Gly Phe Asp Glu Leu
Leu Ala Gln Ala 1400 1405 1410 Arg Gln Arg Ala Leu Glu Ala Gln Ala
His Gln Asp Leu Pro Phe 1415 1420 1425 Glu Gln Leu Val Glu Ala Leu
Gln Pro Glu Arg Asn Ala Ser His 1430 1435 1440 Asn Pro Leu Phe Gln
Val Leu Phe Asn His Gln Ser Glu Ile Arg 1445 1450 1455 Ser Val Thr
Pro Glu Val Gln Leu Glu Asp Leu Arg Leu Glu Gly 1460 1465 1470 Leu
Ala Trp Asp Gly Gln Thr Ala Gln Phe Asp Leu Thr Leu Asp 1475 1480
1485 Ile Gln Glu Asp Glu Asn Gly Ile Trp Ala Ser Phe Asp Tyr Ala
1490 1495 1500 Thr Asp Leu Phe Asp Ala Ser Thr Val Glu Arg Leu Ala
Gly His 1505 1510 1515 Trp Arg Asn Leu Leu Arg Gly Ile Val Ala Asn
Pro Arg Gln Arg 1520 1525 1530 Leu Gly Glu Leu Pro Leu Leu Asp Ala
Pro Glu Arg Arg Gln Thr 1535 1540 1545 Leu Ser Glu Trp Asn Pro Ala
Gln Arg Glu Cys Ala Val Gln Gly 1550 1555 1560 Thr Leu Gln Gln Arg
Phe Glu Glu Gln Ala Arg Gln Arg Pro Gln 1565 1570 1575 Ala Val Ala
Leu Ile Leu Asp Glu Gln Arg Leu Ser Tyr Gly Glu 1580 1585 1590 Leu
Asn Ala Arg Ala Asn Arg Leu Ala His Cys Leu Ile Ala Arg 1595 1600
1605 Gly Val Gly Ala Asp Val Pro Val Gly Leu Ala Leu Glu Arg Ser
1610 1615 1620 Leu Asp Met Leu Val Gly Leu Leu Ala Ile Leu Lys Ala
Gly Gly 1625 1630 1635 Ala Tyr Leu Pro Leu Asp Pro Ala Ala Pro Glu
Glu Arg Leu Ala 1640 1645 1650 His Ile Leu Asp Asp Ser Gly Val Arg
Leu Leu Leu Thr Gln Gly 1655 1660 1665 His Leu Leu Glu Arg Leu Pro
Arg Gln Ala Gly Val Glu Val Leu 1670 1675 1680 Ala Ile Asp Gly Leu
Val Leu Asp Gly Tyr Ala Glu Ser Asp Pro 1685 1690 1695 Leu Pro Thr
Leu Ser Ala Asp Asn Leu Ala Tyr Val Ile Tyr Thr 1700 1705 1710 Ser
Gly Ser Thr Gly Lys Pro Lys Gly Thr Leu Leu Thr His Arg 1715 1720
1725 Asn Ala Leu Arg Leu Phe Ser Ala Thr Glu Ala Trp Phe Gly Phe
1730 1735 1740 Asp Glu Arg Asp Val Trp Thr Leu Phe His Ser Tyr Ala
Phe Asp 1745 1750 1755 Phe Ser Val Trp Glu Ile Phe Gly Ala Leu Leu
Tyr Gly Gly Arg 1760 1765 1770 Leu Val Ile Val Pro Gln Trp Val Ser
Arg Ser Pro Glu Asp Phe 1775
1780 1785 Tyr Arg Leu Leu Cys Arg Glu Gly Val Thr Val Leu Asn Gln
Thr 1790 1795 1800 Pro Ser Ala Phe Lys Gln Leu Met Ala Val Ala Cys
Ser Ala Asp 1805 1810 1815 Met Ala Thr Gln Gln Pro Ala Leu Arg Tyr
Val Ile Phe Gly Gly 1820 1825 1830 Glu Ala Leu Asp Leu Gln Ser Leu
Arg Pro Trp Phe Gln Arg Phe 1835 1840 1845 Gly Asp Arg Gln Pro Gln
Leu Val Asn Met Tyr Gly Ile Thr Glu 1850 1855 1860 Thr Thr Val His
Val Thr Tyr Arg Pro Val Ser Glu Ala Asp Leu 1865 1870 1875 Lys Gly
Gly Leu Val Ser Pro Ile Gly Gly Thr Ile Pro Asp Leu 1880 1885 1890
Ser Trp Tyr Ile Leu Asp Arg Asp Leu Asn Pro Val Pro Arg Gly 1895
1900 1905 Ala Val Gly Glu Leu Tyr Ile Gly Arg Ala Gly Leu Ala Arg
Gly 1910 1915 1920 Tyr Leu Arg Arg Pro Gly Leu Ser Ala Thr Arg Phe
Val Pro Asn 1925 1930 1935 Pro Phe Pro Gly Gly Ala Gly Glu Arg Leu
Tyr Arg Thr Gly Asp 1940 1945 1950 Leu Ala Arg Phe Gln Ala Asp Gly
Asn Ile Glu Tyr Ile Gly Arg 1955 1960 1965 Ile Asp His Gln Val Lys
Val Arg Gly Phe Arg Ile Glu Leu Gly 1970 1975 1980 Glu Ile Glu Ala
Ala Leu Ala Gly Leu Ala Gly Val Arg Asp Ala 1985 1990 1995 Val Val
Leu Ala His Asp Gly Val Gly Gly Thr Gln Leu Val Gly 2000 2005 2010
Tyr Val Val Ala Asp Ser Ala Glu Asp Ala Glu Arg Leu Arg Glu 2015
2020 2025 Ser Leu Arg Glu Ser Leu Lys Arg His Leu Pro Asp Tyr Met
Val 2030 2035 2040 Pro Ala His Leu Met Leu Leu Glu Arg Met Pro Leu
Thr Val Asn 2045 2050 2055 Gly Lys Leu Asp Arg Gln Ala Leu Pro Gln
Pro Asp Ala Ser Leu 2060 2065 2070 Ser Gln Gln Ala Tyr Arg Ala Pro
Gly Ser Glu Leu Glu Gln Arg 2075 2080 2085 Ile Ala Ala Ile Trp Ala
Glu Ile Leu Gly Val Glu Arg Val Gly 2090 2095 2100 Leu Asp Asp Asn
Phe Phe Glu Leu Gly Gly His Ser Leu Leu Leu 2105 2110 2115 Leu Met
Leu Lys Glu Arg Ile Gly Asp Thr Cys Gln Ala Thr Leu 2120 2125 2130
Ser Ile Ser Gln Leu Met Thr His Ala Ser Val Ala Glu Gln Ala 2135
2140 2145 Ala Cys Ile Glu Gly Gln Ala Arg Glu Ser Leu Leu Val Pro
Leu 2150 2155 2160 Asn Gly Arg Arg Glu Gly Ser Pro Leu Phe Met Phe
His Pro Ser 2165 2170 2175 Phe Gly Ser Val His Cys Tyr Lys Thr Leu
Ala Met Ala Leu Arg 2180 2185 2190 Asp Arg His Pro Val Lys Gly Val
Val Cys Arg Ala Leu Leu Gly 2195 2200 2205 Ala Gly Arg Glu Val Pro
Glu Trp Asp Asp Met Val Ala Glu Tyr 2210 2215 2220 Ala Glu Gln Leu
Leu Gln Glu His Pro Glu Gly Val Phe Asn Leu 2225 2230 2235 Ala Gly
Trp Ser Leu Gly Gly Asn Leu Ala Met Asp Val Ala Ala 2240 2245 2250
Arg Leu Glu Gln Arg Gly Arg Gln Val Ala Phe Val Gly Trp Ile 2255
2260 2265 Asp Ala Pro Ala Pro Val Arg Val Glu Ala Phe Trp Asn Glu
Ile 2270 2275 2280 Gly Pro Thr Pro Glu Ala Val Pro Asn Leu Ser Val
Gly Glu Met 2285 2290 2295 Arg Val Glu Leu Leu Gly Val Met Phe Pro
Glu Arg Ala Glu His 2300 2305 2310 Ile Glu Arg Ala Trp Ser Ser Ile
Cys Ser Ala Thr Thr Asp Asp 2315 2320 2325 Glu Gln Arg Trp Thr Arg
Met Ser Asp Trp Ala Glu Ala Glu Ile 2330 2335 2340 Gly Ala Glu Phe
Ala Thr Leu Arg Ser Glu Ile Ala Gln Ser Asn 2345 2350 2355 Glu Leu
Glu Val Ser Trp Glu Leu Lys Gln Ile Leu Asp Glu Arg 2360 2365 2370
Leu Lys Ala Met Asp Tyr Pro Arg Leu Thr Ala Lys Val Ser Leu 2375
2380 2385 Trp Trp Ala Ala Arg Ser Thr Asn Ala Ile Gln Arg Ser Ala
Val 2390 2395 2400 Glu Arg Ser Met Ala Glu Ala Ile Gly Ala Glu Arg
Val Glu Pro 2405 2410 2415 Val Arg Val Leu Asp Thr Arg His Asp Lys
Ile Ile Asp His Pro 2420 2425 2430 Glu Phe Val Gln Ser Phe Arg Ala
Ala Leu Glu Arg Ala Gly Arg 2435 2440 2445 19 3132 DNA Pseudomonas
aeruginosa 19 atgtccgaat tcttcatcaa gcggccgaac ttcgcctggg
tggtggccct gttcatctcc 60 ctggccggcc tgctggtcat ttccaaattg
ccggtagcgc agtaccccaa tgtcgcgccg 120 ccacagatca ccatcaccgc
cacctatccc ggcgcctcgg cgaaggtgct ggtggactcc 180 gtcaccagtg
tgctcgagga gtcgctgaac ggcgccaagg gcctgctcta cttcgagtcg 240
accaacaact ccaacggcac cgccgagatc gtcgtcacct tcgagccggg caccgatccg
300 gacctggccc aggtggacgt gcagaaccgc ctgaagaaag ccgaggcgcg
catgccgcag 360 gcggtgctga cccagggcct gcaggtcgag cagaccagcg
ccggtttcct gctgatctat 420 gcgctcagct acaaggaagg cgctcagcgc
agcgacacca ccgccctcgg cgactacgcc 480 gcgcgcaata tcaacaacga
gctgcggcgc ctgccgggcg tcggcaagct gcaattcttc 540 tcttccgagg
cggccatgcg ggtctggatc gatccgcaga agctggtggg cttcggcctc 600
tccatcgacg acgtgagcaa tgccatccgc gggcagaacg tgcaggtgcc ggccggcgcc
660 ttcggcagcg caccgggcag ttccgcgcag gagctgacgg cgaccctggc
ggtgaagggc 720 accctggacg atccgcagga gttcggccag gtagtgctgc
gcgccaacga ggacggctcg 780 ctggtccggc tcgccgatgt cgcgcgcctg
gaactcggca aggagagcta caacatttcc 840 tcgcgactga acggcacgcc
caccgtgggc ggggctatcc agctgtcgcc cggggccaac 900 gcgatccaga
ccgctaccct ggtgaaacag cgtctcgccg aactgtcggc gttcttcccc 960
gaggacatgc agtacagcgt gccctacgac acctcgcgct tcgtcgacgt ggccatcgag
1020 aaggtgatcc acaccctgat cgaagcgatg gtcctggtgt tcctggtgat
gttcctgttc 1080 ctgcagaacg tccgctacac cctgatcccg tccatcgtgg
tgccggtgtg cctgctgggt 1140 acgctgatgg tgatgtacct gctggggttc
tcggtgaaca tgatgaccat gttcggcatg 1200 gtcctggcga tcggcatcct
ggtggacgac gccatcgtgg tggtggagaa cgtcgagcgg 1260 atcatggcgg
aggaggggat ttccccggcc gaggccacgg tcaaggcgat gaagcaggta 1320
tccggcgcca tcgtcggcat caccctggtg ctctcggcgg tgttcctgcc gctggctttc
1380 atggccggtt cggtgggggt gatctaccag cagttctcgg tgtcgctggc
ggtctcgatc 1440 ctgttctccg gcttcctcgc cctgaccttc accccggcgc
tgtgcgccac gctgctcaag 1500 cccattcccg aagggcacca cgagaagcgc
ggcttcttcg gcgccttcaa ccgtggcttc 1560 gcccgcgtca ccgagcgcta
ttcgctgctc aactcgaagc tggtggcgcg cgccggacgc 1620 ttcatgctgg
tgtacgccgg cctggtggcc atgctcggct acttctacct gcgcctgccg 1680
gaagccttcg tgccggcgga agacctcggc tacatggtgg tcgacgtgca actgccgcct
1740 ggcgcttcgc gcgtgcgcac cgatgccacc ggcgaggagc tcgagcgctt
cctcaagtcc 1800 cgcgaggcgg tggcttcggt gttcctgatc tcgggcttca
gcttctccgg ccagggcgac 1860 aatgccgcgc tggccttccc aaccttcaag
gactggtccg agcgaggcgc cgagcagtcg 1920 gccgccgccg agatcgccgc
gctgaacgag catttcgcgc tgcccgacga tggcacggtc 1980 atggccgtgt
cgccgccacc gatcaacggt ctgggtaact ccggcggctt cgcattgcgc 2040
ctgatggacc gtagcggggt cggccgcgaa gcgctgctgc aggctcgcga tactcttctt
2100 ggcgagatcc agaccaaccc gaaattcctt tacgcgatga tggaaggact
ggccgaagcg 2160 ccgcaactgc gcctgttgat cgaccgggag aaggcccgtg
ccctgggggt gagcttcgag 2220 accatcagcg gcacgctgtc cgctgccttc
ggctcggagg tgatcaacga cttcaccaat 2280 gcggggcgcc aacagcgggt
ggtgatccag gccgaacagg gcaaccggat gaccccggaa 2340 agcgtgctcg
agctatacgt gcctaacgct gctggcaacc tggtaccgct cagcgccttc 2400
gtcagcgtga aatgggaaga gggaccggtg caattggtgc gctataacgg ctacccgtcg
2460 atccgcatcg tcggtgacgc cgcgcccggc ttcagtaccg gcgaagccat
ggcggaaatg 2520 gagcgcctgg cctcgcagct gccggccggc atcggctacg
agtggaccgg cctgtcctat 2580 caggagaagg tctccgccgg gcaggccacc
agcctgttcg ccctcgccat cctggtggtg 2640 ttcctgttgc tggtggcgct
ctacgagagc tggtcgatcc cgctgtcggt gatgctgatc 2700 gtgccgatcg
gcgccatcgg cgcggtgctc gcggtgatgg tcagcggtat gtccaacgac 2760
gtgtatttca aggtcggcct gatcaccatc atcggtcttt cggcgaagaa cgcgatcctc
2820 atcgtcgagt tcgccaagga actctgggag caggggcata gcctgcgcga
cgccgccatc 2880 gaggccgcgc gcctgcgctt ccggccgatc atcatgactt
ccatggcgtt catcctcggc 2940 gtgatacccc tggccctggc cagcggtgcc
ggcgcggcga gccagcgtgc catcggcacc 3000 ggagtgatcg gcgggatgct
cagcgccacc ttcctcggcg tgctgttcgt acctatctgt 3060 ttcgtctggc
tgctgtcgct gctgcgcagc aagccggcac ccatcgaaca ggccgcttcg 3120
gccggggagt ga 3132 20 1043 PRT Pseudomonas aeruginosa 20 Met Ser
Glu Phe Phe Ile Lys Arg Pro Asn Phe Ala Trp Val Val Ala 1 5 10 15
Leu Phe Ile Ser Leu Ala Gly Leu Leu Val Ile Ser Lys Leu Pro Val 20
25 30 Ala Gln Tyr Pro Asn Val Ala Pro Pro Gln Ile Thr Ile Thr Ala
Thr 35 40 45 Tyr Pro Gly Ala Ser Ala Lys Val Leu Val Asp Ser Val
Thr Ser Val 50 55 60 Leu Glu Glu Ser Leu Asn Gly Ala Lys Gly Leu
Leu Tyr Phe Glu Ser 65 70 75 80 Thr Asn Asn Ser Asn Gly Thr Ala Glu
Ile Val Val Thr Phe Glu Pro 85 90 95 Gly Thr Asp Pro Asp Leu Ala
Gln Val Asp Val Gln Asn Arg Leu Lys 100 105 110 Lys Ala Glu Ala Arg
Met Pro Gln Ala Val Leu Thr Gln Gly Leu Gln 115 120 125 Val Glu Gln
Thr Ser Ala Gly Phe Leu Leu Ile Tyr Ala Leu Ser Tyr 130 135 140 Lys
Glu Gly Ala Gln Arg Ser Asp Thr Thr Ala Leu Gly Asp Tyr Ala 145 150
155 160 Ala Arg Asn Ile Asn Asn Glu Leu Arg Arg Leu Pro Gly Val Gly
Lys 165 170 175 Leu Gln Phe Phe Ser Ser Glu Ala Ala Met Arg Val Trp
Ile Asp Pro 180 185 190 Gln Lys Leu Val Gly Phe Gly Leu Ser Ile Asp
Asp Val Ser Asn Ala 195 200 205 Ile Arg Gly Gln Asn Val Gln Val Pro
Ala Gly Ala Phe Gly Ser Ala 210 215 220 Pro Gly Ser Ser Ala Gln Glu
Leu Thr Ala Thr Leu Ala Val Lys Gly 225 230 235 240 Thr Leu Asp Asp
Pro Gln Glu Phe Gly Gln Val Val Leu Arg Ala Asn 245 250 255 Glu Asp
Gly Ser Leu Val Arg Leu Ala Asp Val Ala Arg Leu Glu Leu 260 265 270
Gly Lys Glu Ser Tyr Asn Ile Ser Ser Arg Leu Asn Gly Thr Pro Thr 275
280 285 Val Gly Gly Ala Ile Gln Leu Ser Pro Gly Ala Asn Ala Ile Gln
Thr 290 295 300 Ala Thr Leu Val Lys Gln Arg Leu Ala Glu Leu Ser Ala
Phe Phe Pro 305 310 315 320 Glu Asp Met Gln Tyr Ser Val Pro Tyr Asp
Thr Ser Arg Phe Val Asp 325 330 335 Val Ala Ile Glu Lys Val Ile His
Thr Leu Ile Glu Ala Met Val Leu 340 345 350 Val Phe Leu Val Met Phe
Leu Phe Leu Gln Asn Val Arg Tyr Thr Leu 355 360 365 Ile Pro Ser Ile
Val Val Pro Val Cys Leu Leu Gly Thr Leu Met Val 370 375 380 Met Tyr
Leu Leu Gly Phe Ser Val Asn Met Met Thr Met Phe Gly Met 385 390 395
400 Val Leu Ala Ile Gly Ile Leu Val Asp Asp Ala Ile Val Val Val Glu
405 410 415 Asn Val Glu Arg Ile Met Ala Glu Glu Gly Ile Ser Pro Ala
Glu Ala 420 425 430 Thr Val Lys Ala Met Lys Gln Val Ser Gly Ala Ile
Val Gly Ile Thr 435 440 445 Leu Val Leu Ser Ala Val Phe Leu Pro Leu
Ala Phe Met Ala Gly Ser 450 455 460 Val Gly Val Ile Tyr Gln Gln Phe
Ser Val Ser Leu Ala Val Ser Ile 465 470 475 480 Leu Phe Ser Gly Phe
Leu Ala Leu Thr Phe Thr Pro Ala Leu Cys Ala 485 490 495 Thr Leu Leu
Lys Pro Ile Pro Glu Gly His His Glu Lys Arg Gly Phe 500 505 510 Phe
Gly Ala Phe Asn Arg Gly Phe Ala Arg Val Thr Glu Arg Tyr Ser 515 520
525 Leu Leu Asn Ser Lys Leu Val Ala Arg Ala Gly Arg Phe Met Leu Val
530 535 540 Tyr Ala Gly Leu Val Ala Met Leu Gly Tyr Phe Tyr Leu Arg
Leu Pro 545 550 555 560 Glu Ala Phe Val Pro Ala Glu Asp Leu Gly Tyr
Met Val Val Asp Val 565 570 575 Gln Leu Pro Pro Gly Ala Ser Arg Val
Arg Thr Asp Ala Thr Gly Glu 580 585 590 Glu Leu Glu Arg Phe Leu Lys
Ser Arg Glu Ala Val Ala Ser Val Phe 595 600 605 Leu Ile Ser Gly Phe
Ser Phe Ser Gly Gln Gly Asp Asn Ala Ala Leu 610 615 620 Ala Phe Pro
Thr Phe Lys Asp Trp Ser Glu Arg Gly Ala Glu Gln Ser 625 630 635 640
Ala Ala Ala Glu Ile Ala Ala Leu Asn Glu His Phe Ala Leu Pro Asp 645
650 655 Asp Gly Thr Val Met Ala Val Ser Pro Pro Pro Ile Asn Gly Leu
Gly 660 665 670 Asn Ser Gly Gly Phe Ala Leu Arg Leu Met Asp Arg Ser
Gly Val Gly 675 680 685 Arg Glu Ala Leu Leu Gln Ala Arg Asp Thr Leu
Leu Gly Glu Ile Gln 690 695 700 Thr Asn Pro Lys Phe Leu Tyr Ala Met
Met Glu Gly Leu Ala Glu Ala 705 710 715 720 Pro Gln Leu Arg Leu Leu
Ile Asp Arg Glu Lys Ala Arg Ala Leu Gly 725 730 735 Val Ser Phe Glu
Thr Ile Ser Gly Thr Leu Ser Ala Ala Phe Gly Ser 740 745 750 Glu Val
Ile Asn Asp Phe Thr Asn Ala Gly Arg Gln Gln Arg Val Val 755 760 765
Ile Gln Ala Glu Gln Gly Asn Arg Met Thr Pro Glu Ser Val Leu Glu 770
775 780 Leu Tyr Val Pro Asn Ala Ala Gly Asn Leu Val Pro Leu Ser Ala
Phe 785 790 795 800 Val Ser Val Lys Trp Glu Glu Gly Pro Val Gln Leu
Val Arg Tyr Asn 805 810 815 Gly Tyr Pro Ser Ile Arg Ile Val Gly Asp
Ala Ala Pro Gly Phe Ser 820 825 830 Thr Gly Glu Ala Met Ala Glu Met
Glu Arg Leu Ala Ser Gln Leu Pro 835 840 845 Ala Gly Ile Gly Tyr Glu
Trp Thr Gly Leu Ser Tyr Gln Glu Lys Val 850 855 860 Ser Ala Gly Gln
Ala Thr Ser Leu Phe Ala Leu Ala Ile Leu Val Val 865 870 875 880 Phe
Leu Leu Leu Val Ala Leu Tyr Glu Ser Trp Ser Ile Pro Leu Ser 885 890
895 Val Met Leu Ile Val Pro Ile Gly Ala Ile Gly Ala Val Leu Ala Val
900 905 910 Met Val Ser Gly Met Ser Asn Asp Val Tyr Phe Lys Val Gly
Leu Ile 915 920 925 Thr Ile Ile Gly Leu Ser Ala Lys Asn Ala Ile Leu
Ile Val Glu Phe 930 935 940 Ala Lys Glu Leu Trp Glu Gln Gly His Ser
Leu Arg Asp Ala Ala Ile 945 950 955 960 Glu Ala Ala Arg Leu Arg Phe
Arg Pro Ile Ile Met Thr Ser Met Ala 965 970 975 Phe Ile Leu Gly Val
Ile Pro Leu Ala Leu Ala Ser Gly Ala Gly Ala 980 985 990 Ala Ser Gln
Arg Ala Ile Gly Thr Gly Val Ile Gly Gly Met Leu Ser 995 1000 1005
Ala Thr Phe Leu Gly Val Leu Phe Val Pro Ile Cys Phe Val Trp 1010
1015 1020 Leu Leu Ser Leu Leu Arg Ser Lys Pro Ala Pro Ile Glu Gln
Ala 1025 1030 1035 Ala Ser Ala Gly Glu 1040 21 642 DNA Pseudomonas
aeruginosa 21 atgaacgatg cttctccccg tctgaccgaa cgcggcaggc
aacgccgccg cgccatgctc 60 gacgccgcta cccaggcctt tctcgaacac
ggtttcgaag gcaccaccct ggacatggtg 120 atagaacggg ccggtggttc
acgggggacc ctgtacagct ccttcggcgg caaggagggc 180 ctgttcgccg
cggtgatcgc ccacatgatc ggggaaatct tcgacgacag cgccgatcag 240
ccgcgccccg ccgccacgct gagcgccacc ctcgagcatt tcggccggcg ctttctcacc
300 agcctgctcg atccccgctg ccagagcctc tatcgcctgg tggtggcgga
atccccgcgg 360 tttccggcga tcggcaagtc cttctacgag caggggccgc
agcagagcta tctgctgctc 420 agcgagcgac tggccgcggt cgctcctcac
atggacgagg aaacgctcta cgcggtggcc 480 tgccagtttc tcgagatgct
caaggccgac ctgttcctca aggccctcag cgtggccgac 540 ttccagccga
ccatggcgct gctggaaacc cgcctcaagc tgtcggtgga catcatcgcc 600
tgctacctgg aacacctgtc gcagagcccc gcgcagggct ga 642 22 213 PRT
Pseudomonas aeruginosa 22 Met Asn Asp Ala Ser Pro Arg Leu Thr Glu
Arg Gly Arg Gln Arg Arg 1 5 10 15 Arg Ala Met Leu Asp Ala Ala Thr
Gln Ala Phe Leu Glu His Gly Phe 20 25 30 Glu Gly Thr Thr Leu Asp
Met Val Ile Glu Arg Ala Gly Gly Ser Arg 35 40 45 Gly Thr Leu Tyr
Ser Ser Phe Gly Gly Lys Glu Gly Leu Phe Ala Ala 50 55 60 Val Ile
Ala His Met Ile Gly Glu Ile Phe Asp Asp Ser Ala Asp Gln 65 70 75 80
Pro Arg
Pro Ala Ala Thr Leu Ser Ala Thr Leu Glu His Phe Gly Arg 85 90 95
Arg Phe Leu Thr Ser Leu Leu Asp Pro Arg Cys Gln Ser Leu Tyr Arg 100
105 110 Leu Val Val Ala Glu Ser Pro Arg Phe Pro Ala Ile Gly Lys Ser
Phe 115 120 125 Tyr Glu Gln Gly Pro Gln Gln Ser Tyr Leu Leu Leu Ser
Glu Arg Leu 130 135 140 Ala Ala Val Ala Pro His Met Asp Glu Glu Thr
Leu Tyr Ala Val Ala 145 150 155 160 Cys Gln Phe Leu Glu Met Leu Lys
Ala Asp Leu Phe Leu Lys Ala Leu 165 170 175 Ser Val Ala Asp Phe Gln
Pro Thr Met Ala Leu Leu Glu Thr Arg Leu 180 185 190 Lys Leu Ser Val
Asp Ile Ile Ala Cys Tyr Leu Glu His Leu Ser Gln 195 200 205 Ser Pro
Ala Gln Gly 210 23 1017 DNA Pseudomonas aeruginosa 23 atgtctgatg
atgcccgttt ccagcagctg aattgctggt tggactcttg tttgcccgag 60
ttgttcgttg ccgaaggttg gggggaagtg ccccccgccg aactgatccc ggccagtagc
120 gacgccagct tccgtcgtta tttccgctgg cagggagggg accgcagcct
ggtggtgatg 180 gacgcgccgc cgccccagga agactgccga ccgttcgtca
aggtcgccgg actgctcgcc 240 ggagccggcg tgcatgtgcc gaggattctc
gcccaggacc tggagaacgg tttcctgctg 300 ctcagtgacc tgggccggca
gacctacctc gacgtgcttc atcccgggaa tgccgacgag 360 ctgttcgaac
cggccctgga tgcgctgatc gccttccaga aggtcgatgt cgccggtgtc 420
ctgcctgcct acgacgaagc ggtgctgcgc cgcgagctgc agctgttccc cgactggtac
480 ctggcccgcc acctcggcgt ggagctggag ggcgagacgc tggcccgctg
gaaacggatc 540 tgcgacctgc tggtacgcag cgcgctggag caaccgcggg
tgttcgtcca tcgcgactat 600 atgccgcgca atctgatgct cagcgagccc
aacccgggcg tcctcgactt ccaggacgcc 660 ctgcacggcc cggtcaccta
cgatgtcacc tgcctgtaca aggacgcctt cgtcagttgg 720 ccggagccgc
gcgtgcatgc cgcgctgaac cgttactgga agaaggcgac ctgggccggc 780
atcccgctgc cgccaagctt cgaagacttc ctccgtgcca gcgacctgat gggcgtgcag
840 cgccacctga aggtgattgg catcttcgcc cgtatctgtc accgcgacgg
caagccgcgc 900 tacctgggtg acgtgccgcg cttcttccgt tatctggaaa
ccgccgtggc gcgccgtccc 960 gagctggccg aactgggcga gctgctggcc
tcgctgccgc agggagccga ggcatga 1017 24 338 PRT Pseudomonas
aeruginosa 24 Met Ser Asp Asp Ala Arg Phe Gln Gln Leu Asn Cys Trp
Leu Asp Ser 1 5 10 15 Cys Leu Pro Glu Leu Phe Val Ala Glu Gly Trp
Gly Glu Val Pro Pro 20 25 30 Ala Glu Leu Ile Pro Ala Ser Ser Asp
Ala Ser Phe Arg Arg Tyr Phe 35 40 45 Arg Trp Gln Gly Gly Asp Arg
Ser Leu Val Val Met Asp Ala Pro Pro 50 55 60 Pro Gln Glu Asp Cys
Arg Pro Phe Val Lys Val Ala Gly Leu Leu Ala 65 70 75 80 Gly Ala Gly
Val His Val Pro Arg Ile Leu Ala Gln Asp Leu Glu Asn 85 90 95 Gly
Phe Leu Leu Leu Ser Asp Leu Gly Arg Gln Thr Tyr Leu Asp Val 100 105
110 Leu His Pro Gly Asn Ala Asp Glu Leu Phe Glu Pro Ala Leu Asp Ala
115 120 125 Leu Ile Ala Phe Gln Lys Val Asp Val Ala Gly Val Leu Pro
Ala Tyr 130 135 140 Asp Glu Ala Val Leu Arg Arg Glu Leu Gln Leu Phe
Pro Asp Trp Tyr 145 150 155 160 Leu Ala Arg His Leu Gly Val Glu Leu
Glu Gly Glu Thr Leu Ala Arg 165 170 175 Trp Lys Arg Ile Cys Asp Leu
Leu Val Arg Ser Ala Leu Glu Gln Pro 180 185 190 Arg Val Phe Val His
Arg Asp Tyr Met Pro Arg Asn Leu Met Leu Ser 195 200 205 Glu Pro Asn
Pro Gly Val Leu Asp Phe Gln Asp Ala Leu His Gly Pro 210 215 220 Val
Thr Tyr Asp Val Thr Cys Leu Tyr Lys Asp Ala Phe Val Ser Trp 225 230
235 240 Pro Glu Pro Arg Val His Ala Ala Leu Asn Arg Tyr Trp Lys Lys
Ala 245 250 255 Thr Trp Ala Gly Ile Pro Leu Pro Pro Ser Phe Glu Asp
Phe Leu Arg 260 265 270 Ala Ser Asp Leu Met Gly Val Gln Arg His Leu
Lys Val Ile Gly Ile 275 280 285 Phe Ala Arg Ile Cys His Arg Asp Gly
Lys Pro Arg Tyr Leu Gly Asp 290 295 300 Val Pro Arg Phe Phe Arg Tyr
Leu Glu Thr Ala Val Ala Arg Arg Pro 305 310 315 320 Glu Leu Ala Glu
Leu Gly Glu Leu Leu Ala Ser Leu Pro Gln Gly Ala 325 330 335 Glu Ala
25 3270 DNA Pseudomonas aeruginosa 25 atgagcggat tccaggacca
gagtatcgac gaaggcgtgc gcaagcgcac cgcctaccag 60 aacgatcggc
gtgcacgact ggcattgaac gtcgagcgac aggacggcgg tatcctgcag 120
attccggtgg ccagcgatat gctcggccat gaggagcacg agcgtatcca gcagaacacc
180 ttcctggctg tgatgccgct ggtccgcctg ccaacgctgg gcaaggccgg
ttatggcgac 240 cagctgcccg ccggcgcgct accgcgggcg ggacggatct
acctgttcca ggacggcaag 300 ttgtggcgcg aactggaatg tgatggcaag
ggcaacctgt tcgaagtcga tctcctgcag 360 gggcgcagcc agcgtgcgga
caagcgtccg gccttaggca agacacaagc gctgatcctg 420 gtgccggtgc
tggtcaaggg gcagttcgtg atcccacgct acaccatggc ctatagcgaa 480
actccctggc cttggtcgta catcgactgg ctggaggagg acccgcagcg ggtcaaccgg
540 cgctgccagc agatggcgtc cgcttggaac gcctcggtgg ccaaccagca
ctggaaagcc 600 tccatccatc aacccgcgct ggtcattgat catcacgccc
agggtttgcg acctcgcgac 660 ttcaacgtcg agagcgcgct ggaagacccg
gcggaattca cacctgagtt cgccgccttt 720 cgcgaagagt cgctggtgtg
ccagttgcag cgacgccagc aggaattggc gcccctgctg 780 aagcaggctc
cgccctctgc gctacctact ctggaagccg gagaggacgt actggaaacc 840
ctcaagctgc gtggccatcc caacctcatc gggctgatgc tcgacgactc gctgttcgcc
900 ttgcgccacg ctgcggcgca ggcgcgccac tgcgccgcct acttgcgcag
cctcaatgca 960 ctgctgccgc accgtcccaa cggacgctat gcacaggtgc
tgagcaacat gctcgacggc 1020 ccgctcgcca agctcagggg cgaggtcgat
caggccgaac tggacgaggc gatcttcgcc 1080 gaggagcgac agtcttgccg
aatccacctg acgcagcagg tcgagcatct ggttgccctg 1140 ctggaaggcc
ccttgcaccc ggtgttgcag gactggaccc accagtgcga cgaagccctg 1200
ctggagccct acagcctgat gagcgaggca ctggctgcgc tgaaccagct tcccgaccgc
1260 tgcgacgcac tgtacagcgg taccgcctac cgggcgctgg cggcacatgt
cgagcgggtg 1320 gtcagcacgg ttctgcaggc aagccacccg cttggcgcca
tgctcctggc caaggacgaa 1380 ggacaacttc ccgagccggt tcggcgcctg
caggcgctgc gcgatagccc gcggacgccg 1440 gaccccgatg caatgggcct
cagcacgctg atgctgggag ccagtctgct gggcgaggtc 1500 gaccagccca
gcgccggcaa gagcctcgcc tacttcctcg gcgacctgct ggacgtgttc 1560
ggcgccagcg tagtcgagca actcggccgg ctgtcccagg gcgccaccca gatccagctc
1620 gaccgcttgt tcgcaccgac cttcaatact ctgagcgccc tctcggtgaa
gatgaaaggt 1680 atccgcctgc tgcccgacag tcaggtgccg ctcgacatgg
ttgtcgtcgg cgtgcgcgga 1740 gccggcctgc gcaacggtct gaccgaggtc
gagcgccagg agctgaggcg caagagctat 1800 cggcgcgcca tcgttcagga
cggtgccggc aatcccctgg ccggcaccag tccccgcgac 1860 accggcatga
gtcgcgccaa cctgcgcaac gtcatggtgg tggcggtacc caaggatcac 1920
ccggacctgc ttgcctacac gaaattccgt acgcagttag gcacgttgac ccaggtgatg
1980 gagaacactc gcatcgtgcc gacgatgatg ctggggtttg cgatttataa
cttgaatgtg 2040 caggtgcagg catacagtgg ctttgtagac agtggagaaa
agcacagagg gacgatcggg 2100 gctgtcggtg cagtaatcga tttaacagcc
gctggaggaa gccatgcaaa gctgcttttc 2160 ggaccatcta ctgcaaagta
tctagaaacc ccacgtatat cggtagccca aatatcccct 2220 cgatgggcca
ggaatctaga agttcaaaca ggcagcccta agttagggtt gctacgtggg 2280
cttggtggcg cagccacact attcggtgca ggcatcagtg tatgggatgg ctaccgagct
2340 ttgaggcagg gagatagcga tgcggctgcg gcctacggtg tggccgcagt
gggtgggggc 2400 ctttggggtg cctacgtcct aggatggata gtaaaccctt
atgctttgct ggctggtgcg 2460 gttttggcga tcggaggcac tgtggtcgct
aatctactga ctgacagcga tgcggaaacc 2520 atcgtaaaga aaggcccctt
cggccggcaa ttcgccgagg ctggcctgct cgattcgctg 2580 atgggccagg
accagcgctt cgcccatctg aaagacccgc aaacggccta tcgccaattg 2640
ctgggagtcc tcggccatcc gcgggtcttt gtccatcgcc tggaagactg gcgcaaattg
2700 gcgccggcgg cgcatcgatc tgtcttgcag gaagcggaac ggggtcgcca
agcggtcagc 2760 cgcactgcgc tatcctgcat cgaccccaag ttgcaggcgc
tggaggcaaa cgattgggcc 2820 gtggtgctga gttccccgct cctggccatg
ttcgagaatg gccagaaggc gttccgcctg 2880 gtggcccagg agtttctcag
cagcttgccg atcgatccgg gcaccctgtt cggcgtcaag 2940 cgctaccatc
gggtccccgc gggccccgcc aagctcgaag ccttgccgtt ggatgctgcc 3000
agcgtgctct atgtgctgcc ggccagcctg ccgattccgc agttgtctcc tcgggcccgc
3060 tatagcatgc gcatgaccca gggtttgaag atcagcgcac agttcgaact
caatgccgac 3120 cagcctgagc agcggcttgt cctgcctcaa cccagcccga
agagttggag tgcattcaca 3180 tccgccaatc ggtaccttcc cccggacgac
ttgggccccc atgctgcgcc accttattgg 3240 ttgatagaga acagtgagtt
caacgtatga 3270 26 1089 PRT Pseudomonas aeruginosa 26 Met Ser Gly
Phe Gln Asp Gln Ser Ile Asp Glu Gly Val Arg Lys Arg 1 5 10 15 Thr
Ala Tyr Gln Asn Asp Arg Arg Ala Arg Leu Ala Leu Asn Val Glu 20 25
30 Arg Gln Asp Gly Gly Ile Leu Gln Ile Pro Val Ala Ser Asp Met Leu
35 40 45 Gly His Glu Glu His Glu Arg Ile Gln Gln Asn Thr Phe Leu
Ala Val 50 55 60 Met Pro Leu Val Arg Leu Pro Thr Leu Gly Lys Ala
Gly Tyr Gly Asp 65 70 75 80 Gln Leu Pro Ala Gly Ala Leu Pro Arg Ala
Gly Arg Ile Tyr Leu Phe 85 90 95 Gln Asp Gly Lys Leu Trp Arg Glu
Leu Glu Cys Asp Gly Lys Gly Asn 100 105 110 Leu Phe Glu Val Asp Leu
Leu Gln Gly Arg Ser Gln Arg Ala Asp Lys 115 120 125 Arg Pro Ala Leu
Gly Lys Thr Gln Ala Leu Ile Leu Val Pro Val Leu 130 135 140 Val Lys
Gly Gln Phe Val Ile Pro Arg Tyr Thr Met Ala Tyr Ser Glu 145 150 155
160 Thr Pro Trp Pro Trp Ser Tyr Ile Asp Trp Leu Glu Glu Asp Pro Gln
165 170 175 Arg Val Asn Arg Arg Cys Gln Gln Met Ala Ser Ala Trp Asn
Ala Ser 180 185 190 Val Ala Asn Gln His Trp Lys Ala Ser Ile His Gln
Pro Ala Leu Val 195 200 205 Ile Asp His His Ala Gln Gly Leu Arg Pro
Arg Asp Phe Asn Val Glu 210 215 220 Ser Ala Leu Glu Asp Pro Ala Glu
Phe Thr Pro Glu Phe Ala Ala Phe 225 230 235 240 Arg Glu Glu Ser Leu
Val Cys Gln Leu Gln Arg Arg Gln Gln Glu Leu 245 250 255 Ala Pro Leu
Leu Lys Gln Ala Pro Pro Ser Ala Leu Pro Thr Leu Glu 260 265 270 Ala
Gly Glu Asp Val Leu Glu Thr Leu Lys Leu Arg Gly His Pro Asn 275 280
285 Leu Ile Gly Leu Met Leu Asp Asp Ser Leu Phe Ala Leu Arg His Ala
290 295 300 Ala Ala Gln Ala Arg His Cys Ala Ala Tyr Leu Arg Ser Leu
Asn Ala 305 310 315 320 Leu Leu Pro His Arg Pro Asn Gly Arg Tyr Ala
Gln Val Leu Ser Asn 325 330 335 Met Leu Asp Gly Pro Leu Ala Lys Leu
Arg Gly Glu Val Asp Gln Ala 340 345 350 Glu Leu Asp Glu Ala Ile Phe
Ala Glu Glu Arg Gln Ser Cys Arg Ile 355 360 365 His Leu Thr Gln Gln
Val Glu His Leu Val Ala Leu Leu Glu Gly Pro 370 375 380 Leu His Pro
Val Leu Gln Asp Trp Thr His Gln Cys Asp Glu Ala Leu 385 390 395 400
Leu Glu Pro Tyr Ser Leu Met Ser Glu Ala Leu Ala Ala Leu Asn Gln 405
410 415 Leu Pro Asp Arg Cys Asp Ala Leu Tyr Ser Gly Thr Ala Tyr Arg
Ala 420 425 430 Leu Ala Ala His Val Glu Arg Val Val Ser Thr Val Leu
Gln Ala Ser 435 440 445 His Pro Leu Gly Ala Met Leu Leu Ala Lys Asp
Glu Gly Gln Leu Pro 450 455 460 Glu Pro Val Arg Arg Leu Gln Ala Leu
Arg Asp Ser Pro Arg Thr Pro 465 470 475 480 Asp Pro Asp Ala Met Gly
Leu Ser Thr Leu Met Leu Gly Ala Ser Leu 485 490 495 Leu Gly Glu Val
Asp Gln Pro Ser Ala Gly Lys Ser Leu Ala Tyr Phe 500 505 510 Leu Gly
Asp Leu Leu Asp Val Phe Gly Ala Ser Val Val Glu Gln Leu 515 520 525
Gly Arg Leu Ser Gln Gly Ala Thr Gln Ile Gln Leu Asp Arg Leu Phe 530
535 540 Ala Pro Thr Phe Asn Thr Leu Ser Ala Leu Ser Val Lys Met Lys
Gly 545 550 555 560 Ile Arg Leu Leu Pro Asp Ser Gln Val Pro Leu Asp
Met Val Val Val 565 570 575 Gly Val Arg Gly Ala Gly Leu Arg Asn Gly
Leu Thr Glu Val Glu Arg 580 585 590 Gln Glu Leu Arg Arg Lys Ser Tyr
Arg Arg Ala Ile Val Gln Asp Gly 595 600 605 Ala Gly Asn Pro Leu Ala
Gly Thr Ser Pro Arg Asp Thr Gly Met Ser 610 615 620 Arg Ala Asn Leu
Arg Asn Val Met Val Val Ala Val Pro Lys Asp His 625 630 635 640 Pro
Asp Leu Leu Ala Tyr Thr Lys Phe Arg Thr Gln Leu Gly Thr Leu 645 650
655 Thr Gln Val Met Glu Asn Thr Arg Ile Val Pro Thr Met Met Leu Gly
660 665 670 Phe Ala Ile Tyr Asn Leu Asn Val Gln Val Gln Ala Tyr Ser
Gly Phe 675 680 685 Val Asp Ser Gly Glu Lys His Arg Gly Thr Ile Gly
Ala Val Gly Ala 690 695 700 Val Ile Asp Leu Thr Ala Ala Gly Gly Ser
His Ala Lys Leu Leu Phe 705 710 715 720 Gly Pro Ser Thr Ala Lys Tyr
Leu Glu Thr Pro Arg Ile Ser Val Ala 725 730 735 Gln Ile Ser Pro Arg
Trp Ala Arg Asn Leu Glu Val Gln Thr Gly Ser 740 745 750 Pro Lys Leu
Gly Leu Leu Arg Gly Leu Gly Gly Ala Ala Thr Leu Phe 755 760 765 Gly
Ala Gly Ile Ser Val Trp Asp Gly Tyr Arg Ala Leu Arg Gln Gly 770 775
780 Asp Ser Asp Ala Ala Ala Ala Tyr Gly Val Ala Ala Val Gly Gly Gly
785 790 795 800 Leu Trp Gly Ala Tyr Val Leu Gly Trp Ile Val Asn Pro
Tyr Ala Leu 805 810 815 Leu Ala Gly Ala Val Leu Ala Ile Gly Gly Thr
Val Val Ala Asn Leu 820 825 830 Leu Thr Asp Ser Asp Ala Glu Thr Ile
Val Lys Lys Gly Pro Phe Gly 835 840 845 Arg Gln Phe Ala Glu Ala Gly
Leu Leu Asp Ser Leu Met Gly Gln Asp 850 855 860 Gln Arg Phe Ala His
Leu Lys Asp Pro Gln Thr Ala Tyr Arg Gln Leu 865 870 875 880 Leu Gly
Val Leu Gly His Pro Arg Val Phe Val His Arg Leu Glu Asp 885 890 895
Trp Arg Lys Leu Ala Pro Ala Ala His Arg Ser Val Leu Gln Glu Ala 900
905 910 Glu Arg Gly Arg Gln Ala Val Ser Arg Thr Ala Leu Ser Cys Ile
Asp 915 920 925 Pro Lys Leu Gln Ala Leu Glu Ala Asn Asp Trp Ala Val
Val Leu Ser 930 935 940 Ser Pro Leu Leu Ala Met Phe Glu Asn Gly Gln
Lys Ala Phe Arg Leu 945 950 955 960 Val Ala Gln Glu Phe Leu Ser Ser
Leu Pro Ile Asp Pro Gly Thr Leu 965 970 975 Phe Gly Val Lys Arg Tyr
His Arg Val Pro Ala Gly Pro Ala Lys Leu 980 985 990 Glu Ala Leu Pro
Leu Asp Ala Ala Ser Val Leu Tyr Val Leu Pro Ala 995 1000 1005 Ser
Leu Pro Ile Pro Gln Leu Ser Pro Arg Ala Arg Tyr Ser Met 1010 1015
1020 Arg Met Thr Gln Gly Leu Lys Ile Ser Ala Gln Phe Glu Leu Asn
1025 1030 1035 Ala Asp Gln Pro Glu Gln Arg Leu Val Leu Pro Gln Pro
Ser Pro 1040 1045 1050 Lys Ser Trp Ser Ala Phe Thr Ser Ala Asn Arg
Tyr Leu Pro Pro 1055 1060 1065 Asp Asp Leu Gly Pro His Ala Ala Pro
Pro Tyr Trp Leu Ile Glu 1070 1075 1080 Asn Ser Glu Phe Asn Val 1085
27 756 DNA Pseudomonas aeruginosa 27 atgagcgccg cctgggtccg
gccgttccgc ctgacgccga tgccgcgcct gcgcctggcc 60 tgcttccccc
atgcaggcgg cagcgccagc ttcttccgta gctggagcga acgcctgccg 120
ccagacatcg acctgcttgc cctgcagtac ccgggtcgcg aggaccgctt caacgaggcg
180 ccggccaccc gcctggagga cctcgccgac ggcgccgccc tcgccctgcg
cgatttcgcc 240 gacgcgcccc tggcgctgtt cggccacagt ctcggcgcgg
cgctggccta cgaaaccgcc 300 ctgcgcctgg aaagcgccgg cgcgccgctg
cgccacctgt tcgtctccgc ccatccggca 360 ccgcaccggc aacgcggcgg
cgcgttgcac cgcggcgacg aggcggcgct gctggaggac 420 gtccgccgcc
agggtggcgc cagcgagcta ctcgaggacg ccgacctgcg cgcgctgttc 480
ctgccgatcc tgcgcgccga ctaccaggcg atcgagacct accgacgggc gcagcccatc
540 gccctggcct gcgccctcga cgtcctcctc ggcgagcacg acgaggaagt
cagcgccgcc 600 gaggcgcagg cctggagcga cgccagccgg actcccgcca
ggctgcggcg ctttcctggc 660 ggccacttct acctgagcga ggggcgcgac
gcggtgatcg agcacctgct gcgccgcctc 720 gcacatcccg acgccctttc
ccgagaggtt gcatga 756 28 251 PRT Pseudomonas aeruginosa 28 Met Ser
Ala Ala Trp Val Arg Pro Phe Arg Leu Thr Pro Met Pro Arg 1 5 10 15
Leu Arg Leu Ala Cys Phe Pro His Ala Gly Gly Ser Ala Ser Phe Phe
20
25 30 Arg Ser Trp Ser Glu Arg Leu Pro Pro Asp Ile Asp Leu Leu Ala
Leu 35 40 45 Gln Tyr Pro Gly Arg Glu Asp Arg Phe Asn Glu Ala Pro
Ala Thr Arg 50 55 60 Leu Glu Asp Leu Ala Asp Gly Ala Ala Leu Ala
Leu Arg Asp Phe Ala 65 70 75 80 Asp Ala Pro Leu Ala Leu Phe Gly His
Ser Leu Gly Ala Ala Leu Ala 85 90 95 Tyr Glu Thr Ala Leu Arg Leu
Glu Ser Ala Gly Ala Pro Leu Arg His 100 105 110 Leu Phe Val Ser Ala
His Pro Ala Pro His Arg Gln Arg Gly Gly Ala 115 120 125 Leu His Arg
Gly Asp Glu Ala Ala Leu Leu Glu Asp Val Arg Arg Gln 130 135 140 Gly
Gly Ala Ser Glu Leu Leu Glu Asp Ala Asp Leu Arg Ala Leu Phe 145 150
155 160 Leu Pro Ile Leu Arg Ala Asp Tyr Gln Ala Ile Glu Thr Tyr Arg
Arg 165 170 175 Ala Gln Pro Ile Ala Leu Ala Cys Ala Leu Asp Val Leu
Leu Gly Glu 180 185 190 His Asp Glu Glu Val Ser Ala Ala Glu Ala Gln
Ala Trp Ser Asp Ala 195 200 205 Ser Arg Thr Pro Ala Arg Leu Arg Arg
Phe Pro Gly Gly His Phe Tyr 210 215 220 Leu Ser Glu Gly Arg Asp Ala
Val Ile Glu His Leu Leu Arg Arg Leu 225 230 235 240 Ala His Pro Asp
Ala Leu Ser Arg Glu Val Ala 245 250 29 4317 DNA Pseudomonas
aeruginosa 29 atggatctgc cccccgattc ccgtaccgcc ctgcgcgact
ggctgaccga gcagctcgcc 60 gacctgctcg gcgaaccgct tgctgacgtg
cgcgccctgg cggacgacga cgacctgctg 120 ggctgcggcc tcgactcgat
ccgcctgatg tacctgcagg aacgcctgcg cgcgcgtggc 180 tcgacgctgg
acttcgccca gttggcgcag cgcccctgcc tgggggcctg gctcgacctg 240
ctggcctgcg cggaccggct gtccgccccg gcaacggtcg cgctgccgac ggcgcaggat
300 cgcgatcagc cgttcgagct gtcttccgtg cagcaggcct actggctggg
acgtggcgcc 360 ggcgaggtgc tgggcaacgt cagctgccat gcctttctgg
aattccgcac gcgggatgtc 420 gacccgcagc gcctggccgc ggcggcggag
tgcgtgcgtc aacgccaccc gatgttgcgg 480 gcgcgcttcc tcgacggtcg
ccagcagatc cttccgacgc cgccgctgtc ctgcttcgac 540 ctgcaggact
ggcgcacctt acaggtggac gaggccgagc gcgactggca ggcgctgcgc 600
gactggcgcg cccatgaatg cctggcggtg gagcgcggcc aggtgttcct gctcgggctg
660 gtgcgcatgc cgggcggcga ggatcgcctc tggctgagtc tcgacctgct
tgccgccgat 720 gtcgaaagcc tgcgcctgct gctggccgaa ctgggcgttg
cctacctggc gccggagcgc 780 ctggcggagc cgccggcgct gcatttcgcc
gactacctgg cgcaccgtgc ggcgcaacgc 840 gccgaggccg cggcgcgggc
ccgcgactac tggctggaac gcctgccgcg cttgccggac 900 gcgccggccc
tgccgttggc ctgcgcgccg gaaagcatcc gccagccgcg cacccggcgc 960
ctggcattcc agctttccgc cggcgagagc cggcgcctgg agcgtcttgc cgcgcagcat
1020 ggcgtgacct tgtccagcgt gttcggctgc gccttcgcgc tggtcctggc
gcgctggagc 1080 gaaagcgcgg aatttctcct caacgtgccg ttgttcgatc
ggcatgccga cgacccgcgt 1140 atcggcgagg tgatcgccga cttcaccacc
ctgttgctgc tggagtgccg gatgcaggcc 1200 ggggtgtcct tcgccgaggc
ggtgaagagc ttccagcgca acctccacgg agccatcgac 1260 cacgccgcat
tccccgccct ggaggtgctc cgcgaggcgc gccggcaggg ccagccacgc 1320
tcggcgccgg tggtgttcgc cagcaacctg ggcgaggagg gcttcgtccc ggcggccttc
1380 cgcgacgctt tcggcgatct ccacgacatg ctctcgcaga ccccgcaggt
ctggctcgac 1440 caccagctct accgggtggg cgacggtatc ctgctggcct
gggatagcgt cgtcggcctg 1500 ttccccgaag gtctgccgga aaccatgttc
gaagcctacg tggggctgct ccagcgtctc 1560 tgcgacagcg cctgggggca
gcccgccgat ctgccgttgc cctgggcgca gcaggcgcgc 1620 cgggccctgc
tcaacggcca gccggcatgc gccacggcgc gcaccctgca tcgcgacttc 1680
ttccttcgcg ccgccgaggc gccggatgcc gacgcgctgc tctatcgcga ccaacgtgtc
1740 acccgcggcg aactggccga gcgtgcgctg cgcatcgccg gcggcctgcg
cgaagccggg 1800 gtgcgccctg gcgacgcggt cgaggtcagc ctgccgcgcg
gaccgcagca ggtcgcggcg 1860 gtattcggcg tgctcgccgc aggcgcctgc
tacgtgccgc tggacatcga ccagccgccc 1920 gcacggcggc gcctgatcga
agaggccgcc ggggtatgcc tggcgatcac cgaggaggac 1980 gatccgcagg
ccttgccgcc gcgcctggat gtccagcgcc tgctgcgcgg cccggcgctg 2040
gccgcccccg tgccgctggc gccgcaggcg agtgcctatg tgatctacac ctcgggctcc
2100 accggggtgc ccaagggcgt cgaggtcagc cacgcggcgg cgatcaatac
catcgacgcg 2160 ctgctcgacc tgctgcgggt gaacgcatcg gatcgcttgc
tggcggtctc cgcgctggac 2220 ttcgatctgt cggtcttcga cctgttcggc
ggcctcggcg ccggtgccag cctggtcctg 2280 ccggcccagg aacaggcgcg
cgatgccgct gcctgggcgg aggctatcca gcggcatgcg 2340 gtgagcctgt
ggaactcggc gccggccttg ctggagatgg ccctcagcct gccggcgagc 2400
caggccgact atcgcagtct gcgggcggtg ctgctgtccg gcgactgggt ggccctggac
2460 ctgcccggcc gcctgcgccc acgttgtgcc gaaggctgcc gcctgcatgt
gctgggtggc 2520 gctaccgaag cgggcatctg gtcgaacctg cagagcgtcg
atacggtgcc gccgcactgg 2580 cgttcgattc cctacggccg gccattgccg
ggacaggcct accgggtggt cgacacccac 2640 gggcgcgacg tgccggacct
ggtggtcggc gagctgtgga tcggcggcgc cagcctggcc 2700 cgcggctatc
gcaacgatcc cgaactcagc gcccggcgtt tcgtccacga tgcccagggc 2760
cgctggtatc gcaccggcga tcgcggtcgc tactggggcg acggtaccct ggaattcctc
2820 ggtcgggtcg accagcaggt gaaagtgcgc ggccagcgca tcgagttggg
cgaggtggag 2880 gccgcgctgt gcgcccaggc tggcgtggag agcgcctgcg
cggcggtgct cggcggtggc 2940 gtggcgagcc tcggcgcggt gctggtaccg
cgcctggcgc cacgggccga aggctccatg 3000 gatctaccgg ccgcacagcc
cttcgccggc ctggcagagg ccgaggcggt actcacccgg 3060 gaaatcctcg
gcgcgctgct ggaggcgccg ctggagctag acgacggttt gcgccggcgc 3120
tggctggact ggctagcgga ctccgccgcc agcgcgctgc cgtcgctcga cgaggcgttg
3180 cgccggctcg gctggcaggc cgcggggctg accgcgatgg gcaacgctct
gcgcggcctg 3240 ctcgccggcg aacaggcgcc ggccgcgctg ctcctcgatc
cctggctggc gccgcaggcg 3300 gtggccgcgc gcctgccgga cggccgcgag
gccctggcgc gcctgctcga agcgctgccg 3360 acgccggctg ccggcgaacg
cctgcgggtg gcggtgctgg atacccgcgc cgggctctgg 3420 ctcgaccagg
gcatggcctc gctgttgcgc ccagggctgg aactgaccct cttcgaacgc 3480
agccgcgtcc tcctcgacgc cgccgccacc cgcttgccgg aacggatcgt ggtgcaggcg
3540 ctggacgacg gcctgctacc tgccgagcac ctcggtcgct acgaccgggt
gatcagcttc 3600 gccgcgctgc acgcctacga ggccagccgc gaaggcctgg
cgctggcggc ggcgctgctg 3660 cgcccgcagg gccgcctgtt gctggtggac
ctgctatgcg agtcgccact ggcgctgctc 3720 ggtgcggcct tgctcgacga
ccggccgctg cgcctggcgg agctgccgag cctgttggcc 3780 gatctcgccg
ctgcgggact ggcgccgcgt tgcctgtggc gcagcgagcg gatcgccctg 3840
gtcgaggcgc tggcaccggg actcgggctc gacgccgccg cgctccaggc cggcctggag
3900 caacgcctgc cccaggcgat gcggcccgaa cgcctgtggt gcctgccaag
cctgccgttg 3960 aacggcaatg gcaaggtcga tcgtcgccgc ctggccgaga
gcatgacccg cgcactcggc 4020 gagtgtcgtc acgagccctc ggcggaggag
ccgctggaag cccatgagca agcgctggcc 4080 gagtgctggg aagcggttct
caaacgcccg gtgcgtcgtc gcgaggcgag cttcttcagc 4140 ctcggcggcg
acagcctgct ggcgacccgc ctgctggccg gcatacgtga gcgtttcggc 4200
gtacgcctgg gcatggccga cttctatcgc cagccgaccc tggccggtct tgcccgccac
4260 ttgcaggtgc agaccgtcga aatcgaggaa acccaactgg aagagggcgt gctatga
4317 30 1438 PRT Pseudomonas aeruginosa 30 Met Asp Leu Pro Pro Asp
Ser Arg Thr Ala Leu Arg Asp Trp Leu Thr 1 5 10 15 Glu Gln Leu Ala
Asp Leu Leu Gly Glu Pro Leu Ala Asp Val Arg Ala 20 25 30 Leu Ala
Asp Asp Asp Asp Leu Leu Gly Cys Gly Leu Asp Ser Ile Arg 35 40 45
Leu Met Tyr Leu Gln Glu Arg Leu Arg Ala Arg Gly Ser Thr Leu Asp 50
55 60 Phe Ala Gln Leu Ala Gln Arg Pro Cys Leu Gly Ala Trp Leu Asp
Leu 65 70 75 80 Leu Ala Cys Ala Asp Arg Leu Ser Ala Pro Ala Thr Val
Ala Leu Pro 85 90 95 Thr Ala Gln Asp Arg Asp Gln Pro Phe Glu Leu
Ser Ser Val Gln Gln 100 105 110 Ala Tyr Trp Leu Gly Arg Gly Ala Gly
Glu Val Leu Gly Asn Val Ser 115 120 125 Cys His Ala Phe Leu Glu Phe
Arg Thr Arg Asp Val Asp Pro Gln Arg 130 135 140 Leu Ala Ala Ala Ala
Glu Cys Val Arg Gln Arg His Pro Met Leu Arg 145 150 155 160 Ala Arg
Phe Leu Asp Gly Arg Gln Gln Ile Leu Pro Thr Pro Pro Leu 165 170 175
Ser Cys Phe Asp Leu Gln Asp Trp Arg Thr Leu Gln Val Asp Glu Ala 180
185 190 Glu Arg Asp Trp Gln Ala Leu Arg Asp Trp Arg Ala His Glu Cys
Leu 195 200 205 Ala Val Glu Arg Gly Gln Val Phe Leu Leu Gly Leu Val
Arg Met Pro 210 215 220 Gly Gly Glu Asp Arg Leu Trp Leu Ser Leu Asp
Leu Leu Ala Ala Asp 225 230 235 240 Val Glu Ser Leu Arg Leu Leu Leu
Ala Glu Leu Gly Val Ala Tyr Leu 245 250 255 Ala Pro Glu Arg Leu Ala
Glu Pro Pro Ala Leu His Phe Ala Asp Tyr 260 265 270 Leu Ala His Arg
Ala Ala Gln Arg Ala Glu Ala Ala Ala Arg Ala Arg 275 280 285 Asp Tyr
Trp Leu Glu Arg Leu Pro Arg Leu Pro Asp Ala Pro Ala Leu 290 295 300
Pro Leu Ala Cys Ala Pro Glu Ser Ile Arg Gln Pro Arg Thr Arg Arg 305
310 315 320 Leu Ala Phe Gln Leu Ser Ala Gly Glu Ser Arg Arg Leu Glu
Arg Leu 325 330 335 Ala Ala Gln His Gly Val Thr Leu Ser Ser Val Phe
Gly Cys Ala Phe 340 345 350 Ala Leu Val Leu Ala Arg Trp Ser Glu Ser
Ala Glu Phe Leu Leu Asn 355 360 365 Val Pro Leu Phe Asp Arg His Ala
Asp Asp Pro Arg Ile Gly Glu Val 370 375 380 Ile Ala Asp Phe Thr Thr
Leu Leu Leu Leu Glu Cys Arg Met Gln Ala 385 390 395 400 Gly Val Ser
Phe Ala Glu Ala Val Lys Ser Phe Gln Arg Asn Leu His 405 410 415 Gly
Ala Ile Asp His Ala Ala Phe Pro Ala Leu Glu Val Leu Arg Glu 420 425
430 Ala Arg Arg Gln Gly Gln Pro Arg Ser Ala Pro Val Val Phe Ala Ser
435 440 445 Asn Leu Gly Glu Glu Gly Phe Val Pro Ala Ala Phe Arg Asp
Ala Phe 450 455 460 Gly Asp Leu His Asp Met Leu Ser Gln Thr Pro Gln
Val Trp Leu Asp 465 470 475 480 His Gln Leu Tyr Arg Val Gly Asp Gly
Ile Leu Leu Ala Trp Asp Ser 485 490 495 Val Val Gly Leu Phe Pro Glu
Gly Leu Pro Glu Thr Met Phe Glu Ala 500 505 510 Tyr Val Gly Leu Leu
Gln Arg Leu Cys Asp Ser Ala Trp Gly Gln Pro 515 520 525 Ala Asp Leu
Pro Leu Pro Trp Ala Gln Gln Ala Arg Arg Ala Leu Leu 530 535 540 Asn
Gly Gln Pro Ala Cys Ala Thr Ala Arg Thr Leu His Arg Asp Phe 545 550
555 560 Phe Leu Arg Ala Ala Glu Ala Pro Asp Ala Asp Ala Leu Leu Tyr
Arg 565 570 575 Asp Gln Arg Val Thr Arg Gly Glu Leu Ala Glu Arg Ala
Leu Arg Ile 580 585 590 Ala Gly Gly Leu Arg Glu Ala Gly Val Arg Pro
Gly Asp Ala Val Glu 595 600 605 Val Ser Leu Pro Arg Gly Pro Gln Gln
Val Ala Ala Val Phe Gly Val 610 615 620 Leu Ala Ala Gly Ala Cys Tyr
Val Pro Leu Asp Ile Asp Gln Pro Pro 625 630 635 640 Ala Arg Arg Arg
Leu Ile Glu Glu Ala Ala Gly Val Cys Leu Ala Ile 645 650 655 Thr Glu
Glu Asp Asp Pro Gln Ala Leu Pro Pro Arg Leu Asp Val Gln 660 665 670
Arg Leu Leu Arg Gly Pro Ala Leu Ala Ala Pro Val Pro Leu Ala Pro 675
680 685 Gln Ala Ser Ala Tyr Val Ile Tyr Thr Ser Gly Ser Thr Gly Val
Pro 690 695 700 Lys Gly Val Glu Val Ser His Ala Ala Ala Ile Asn Thr
Ile Asp Ala 705 710 715 720 Leu Leu Asp Leu Leu Arg Val Asn Ala Ser
Asp Arg Leu Leu Ala Val 725 730 735 Ser Ala Leu Asp Phe Asp Leu Ser
Val Phe Asp Leu Phe Gly Gly Leu 740 745 750 Gly Ala Gly Ala Ser Leu
Val Leu Pro Ala Gln Glu Gln Ala Arg Asp 755 760 765 Ala Ala Ala Trp
Ala Glu Ala Ile Gln Arg His Ala Val Ser Leu Trp 770 775 780 Asn Ser
Ala Pro Ala Leu Leu Glu Met Ala Leu Ser Leu Pro Ala Ser 785 790 795
800 Gln Ala Asp Tyr Arg Ser Leu Arg Ala Val Leu Leu Ser Gly Asp Trp
805 810 815 Val Ala Leu Asp Leu Pro Gly Arg Leu Arg Pro Arg Cys Ala
Glu Gly 820 825 830 Cys Arg Leu His Val Leu Gly Gly Ala Thr Glu Ala
Gly Ile Trp Ser 835 840 845 Asn Leu Gln Ser Val Asp Thr Val Pro Pro
His Trp Arg Ser Ile Pro 850 855 860 Tyr Gly Arg Pro Leu Pro Gly Gln
Ala Tyr Arg Val Val Asp Thr His 865 870 875 880 Gly Arg Asp Val Pro
Asp Leu Val Val Gly Glu Leu Trp Ile Gly Gly 885 890 895 Ala Ser Leu
Ala Arg Gly Tyr Arg Asn Asp Pro Glu Leu Ser Ala Arg 900 905 910 Arg
Phe Val His Asp Ala Gln Gly Arg Trp Tyr Arg Thr Gly Asp Arg 915 920
925 Gly Arg Tyr Trp Gly Asp Gly Thr Leu Glu Phe Leu Gly Arg Val Asp
930 935 940 Gln Gln Val Lys Val Arg Gly Gln Arg Ile Glu Leu Gly Glu
Val Glu 945 950 955 960 Ala Ala Leu Cys Ala Gln Ala Gly Val Glu Ser
Ala Cys Ala Ala Val 965 970 975 Leu Gly Gly Gly Val Ala Ser Leu Gly
Ala Val Leu Val Pro Arg Leu 980 985 990 Ala Pro Arg Ala Glu Gly Ser
Met Asp Leu Pro Ala Ala Gln Pro Phe 995 1000 1005 Ala Gly Leu Ala
Glu Ala Glu Ala Val Leu Thr Arg Glu Ile Leu 1010 1015 1020 Gly Ala
Leu Leu Glu Ala Pro Leu Glu Leu Asp Asp Gly Leu Arg 1025 1030 1035
Arg Arg Trp Leu Asp Trp Leu Ala Asp Ser Ala Ala Ser Ala Leu 1040
1045 1050 Pro Ser Leu Asp Glu Ala Leu Arg Arg Leu Gly Trp Gln Ala
Ala 1055 1060 1065 Gly Leu Thr Ala Met Gly Asn Ala Leu Arg Gly Leu
Leu Ala Gly 1070 1075 1080 Glu Gln Ala Pro Ala Ala Leu Leu Leu Asp
Pro Trp Leu Ala Pro 1085 1090 1095 Gln Ala Val Ala Ala Arg Leu Pro
Asp Gly Arg Glu Ala Leu Ala 1100 1105 1110 Arg Leu Leu Glu Ala Leu
Pro Thr Pro Ala Ala Gly Glu Arg Leu 1115 1120 1125 Arg Val Ala Val
Leu Asp Thr Arg Ala Gly Leu Trp Leu Asp Gln 1130 1135 1140 Gly Met
Ala Ser Leu Leu Arg Pro Gly Leu Glu Leu Thr Leu Phe 1145 1150 1155
Glu Arg Ser Arg Val Leu Leu Asp Ala Ala Ala Thr Arg Leu Pro 1160
1165 1170 Glu Arg Ile Val Val Gln Ala Leu Asp Asp Gly Leu Leu Pro
Ala 1175 1180 1185 Glu His Leu Gly Arg Tyr Asp Arg Val Ile Ser Phe
Ala Ala Leu 1190 1195 1200 His Ala Tyr Glu Ala Ser Arg Glu Gly Leu
Ala Leu Ala Ala Ala 1205 1210 1215 Leu Leu Arg Pro Gln Gly Arg Leu
Leu Leu Val Asp Leu Leu Cys 1220 1225 1230 Glu Ser Pro Leu Ala Leu
Leu Gly Ala Ala Leu Leu Asp Asp Arg 1235 1240 1245 Pro Leu Arg Leu
Ala Glu Leu Pro Ser Leu Leu Ala Asp Leu Ala 1250 1255 1260 Ala Ala
Gly Leu Ala Pro Arg Cys Leu Trp Arg Ser Glu Arg Ile 1265 1270 1275
Ala Leu Val Glu Ala Leu Ala Pro Gly Leu Gly Leu Asp Ala Ala 1280
1285 1290 Ala Leu Gln Ala Gly Leu Glu Gln Arg Leu Pro Gln Ala Met
Arg 1295 1300 1305 Pro Glu Arg Leu Trp Cys Leu Pro Ser Leu Pro Leu
Asn Gly Asn 1310 1315 1320 Gly Lys Val Asp Arg Arg Arg Leu Ala Glu
Ser Met Thr Arg Ala 1325 1330 1335 Leu Gly Glu Cys Arg His Glu Pro
Ser Ala Glu Glu Pro Leu Glu 1340 1345 1350 Ala His Glu Gln Ala Leu
Ala Glu Cys Trp Glu Ala Val Leu Lys 1355 1360 1365 Arg Pro Val Arg
Arg Arg Glu Ala Ser Phe Phe Ser Leu Gly Gly 1370 1375 1380 Asp Ser
Leu Leu Ala Thr Arg Leu Leu Ala Gly Ile Arg Glu Arg 1385 1390 1395
Phe Gly Val Arg Leu Gly Met Ala Asp Phe Tyr Arg Gln Pro Thr 1400
1405 1410 Leu Ala Gly Leu Ala Arg His Leu Gln Val Gln Thr Val Glu
Ile 1415 1420 1425 Glu Glu Thr Gln Leu Glu Glu Gly Val Leu 1430
1435 31 5430 DNA Pseudomonas aeruginosa 31 atgagcctcg gcgaactgct
ggaaacctgc cgcagccggc gcatcgaact ctggagcgag 60 gcgggccgcc
tgcgctatcg cgccccccag ggtgccctcg acgccggcct cgccgagcgc 120
ctgcgggccg agcgcgaggc cctgctggaa cacctggaag gcggccctgg ctggcgcgcc
180 gaacccgaca tggcccacca gcgcttcccg ctgaccccgg tgcaggccgc
ctacgtgctg 240 ggccgccagg cggccttcga ctacggcggt aacgcctgcc
agctgtacgc cgagtacgac 300 tggccggccg acaccgatcc ggcgcgcctg
gaggcggcct ggaacgccat ggtcgagcgc 360 cacccgatgc tgcgcgcggt
gatcgaggac aacgcctggc agcgcgtgct gcccgaggtg 420 ccctggcagc
ggctgaccgt gcatgcctgc gcggggctcg acgaggccgc tttccaggcg 480
cacctggagc gggtccgcga acgcctcgac cacgcctgcg cggcgctcga ccagtggccg
540 gtcctgcgcc ccgagctgag tatcggccgg gatgcctgcg tactgcactg
ctcggtggat 600 ttcaccctgg tcgactacgc cagcctgcaa ttgctgcttg
gcgaatggcg ccgccgctat 660 ctcgatccgc aatggacggc ggaaccgctg
gaggcgacct tccgcgacta tgtcggcgtc 720 gagcagcgcc gacgccagtc
gccagcctgg cagcgcgacc gcgactggtg gctggcgcgt 780 ctcgacgcgc
taccggggcg tcccgacctg ccgctgcggg tgcagccgga cacccggtcc 840
acgcgcttcc ggcacttcca cgcgcgcctc gacgaggccg cctggcaggc gctcggcgcg
900 cgcgccggcg aacacggcct gagcgctgcc ggcgtggcct tggcggcctt
cgccgagacc 960 atcggtcgct ggagccaggc accggcgttc tgtctcaacc
tgacggtact caaccggccg 1020 ccgctgcatc cgcagctggc gcaggtgctc
ggtgacttca ccgcgctcag cctgctggca 1080 gtggacagcc gccacggcga
cagtttcgtc gagcgtgccc gacgcatcgg cgagcagatg 1140 ttcgacgacc
tcgaccaccc gaccttcagc ggcgtcgacc tgctgcgcga actggcgcgc 1200
cggcgtggtc gcggcgccga tctgatgccg gtggtgttca ccagtggcat cggcagcgtg
1260 cagcgcctgc tcggcgatgg cgaggcgccg cgcgcgccac gctacatgat
cagccagacc 1320 ccgcaggtct ggctggactg ccaggtcacc gaccagttcg
gcggcctgga gatcggctgg 1380 gacgtacgcc tcgggttgtt ccccgagggc
caggcggaag ccatgttcga cgacttcgtc 1440 gggctgctcc ggcgcctggc
gcagagcccg cgcgcctgga ccgacggcga tgccacggaa 1500 cccgtcgagg
cgccgccgca ggcgttgccc ggtagtgccc ggagcatcgc cgccggtttc 1560
gccgagcgtg ccctgctgac ccccgacgcc acggcgatcc acgatgccgc cggcagctac
1620 agctaccgcc aggtcgccca gcacgccagc gccctgcgcc gcgtcctgga
agcgcacggc 1680 gcgggccgtg gccggcgggt cgcggtgatg ctgccgaaaa
gcgccgcgca attggtcgcg 1740 gtgatcggca tcctccaggc cggcgccgcc
tatgtgccgg tggacatccg ccagcctccg 1800 ctgcggcgcc aggcgatcct
cgccagcgcc gaagtggtcg cgctggtctg cctggaaagc 1860 gatgtcccgg
acgtcggctg cgcctgcgtg gccatcgacc ggctggccgc cgacagcgcc 1920
tggccgccac cgcccgcggc ggaggtggcg gcggacgacc tcgcctacgt gatctacacc
1980 tccggctcca ccggcacgcc aaagggcgtg atgctcagcc atgcggcggt
gagcaacacg 2040 ctgctcgaca tcaaccagcg ctacggcgtc gacgccaacg
accgcgtcct cggcctcgcc 2100 gagctgagct tcgacctctc ggtctacgac
ttcttcggcg ccaccgcggc gggggcccag 2160 gtggtcctcc cggacccggc
gcgcggcagc gatccatcgc actgggcgga actgctggaa 2220 cgccacgcca
tcaccctgtg gaactcggtg ccggcccaag gccagatgct catcgattac 2280
ctggagagcg agccgcaacg tcacctgccg ggaccgcgct gcgtgctctg gtccggtgac
2340 tggattccgg tcagcctgcc gacccgctgg tggcggcgct ggccggacag
cgcgctgttc 2400 agcctgggcg gcgccaccga ggcggcgatc tggtcgatcg
agcagccgat ccgcccgcag 2460 cacaccgagc tggccagcat cccttatggc
cgtgccctgc gcgggcagag cgtggaagtc 2520 ctggatgccc gcgggcggcg
ctgcccgccg ggcgtgcgcg gcgagatcca tatcggcggg 2580 gtgggcctgg
cgctcggcta cgccggcgat ccgcagcgca ccgccgaacg cttcgtccgt 2640
caccccgatg gccgtcgcct gtatcgcacc ggcgacctcg gccgctacct ggccgacggc
2700 agcatcgagt tcctcggccg cgaggacgac caggtgaaga ttcgcggcca
ccgcatcgaa 2760 ctggccgaac tggacgccgc gctgtgcgct catccgcagg
tcaacctggc ggccaccgtg 2820 gtgctcggcg agacccacga gcgcagcctg
gccagcttcg tcaccctgca tgcgccggtg 2880 gaggctggcg aggatccgcg
tacggcgctc gacgcggtgc gccagcgggc ggcccaggcc 2940 ttgcgccgcg
actggggcag cgaggagggc atcgccgcgg cggtggccgc actcgaccgt 3000
gcctgcctcg cctcgttggc cgcctggctg gccggcagcg gtctgttcgc cagtgcgacg
3060 ccgctggact tagccaccct gtgccagcgc ctgggtatcg ccgaggcgcg
ccagcgcctg 3120 ctgcgccact ggttgcgcca actggaggag ggcggctacc
tgcgcgccga gggcgagggc 3180 tggctgggct gcgccgagcg tcccgcgcag
agtccggagg acgcctggac ggcgttcgcc 3240 ggctgcgcgc cggcggcgct
ctggccggcc gagctggtcg cctacctgcg tgacagcgcg 3300 caatccctcg
gcgagcaact ggccgggcgg atcagcccgg cggcgctgat gttcccgcag 3360
ggctcggcgc gcatcgccga ggccatgtac agccagggcc tgcatgccca ggcgctgcac
3420 gaggccatgg ccgaggccat cgccgccatc gtcgagcgcc agccgcaacg
gcgctggcgc 3480 ctgctggagc ttggcgccgg caccgccgcc gccagccgca
cggtgatcgc ccggttggcg 3540 ccgctggtgc agcgaggggc ggaggtggac
tacctgttca ccgacgtttc cagctacttc 3600 ctcgccgccg cccgcgagcg
cttcgccgac cagccgtggg tacgcttcgg ccgcttcgac 3660 atgaacggcg
atcttctcga ccagggcgtg gcgccgcact cggtggatat cctgctcagc 3720
tccggggcct tgaacaacgc gctggacacc ccggcgctgc tggccggcct gcgcgagttg
3780 ctgagcgccg acgcctggct ggtgatccag gaactgacgc gcgagcacaa
cgagatcagc 3840 gtcagccaga gcctgatgat ggaaaacccg cgcgacctcc
gcgacgagcg ccgccaactg 3900 ttcgtccaca ccgggcaatg gctggagtgg
ctggcggcac agggtggcga cctggcttgt 3960 ggggtggtgc cgccgggcag
cgctctcgac ctgcttggct acgatgtcct gctggctcgc 4020 tgcaagaccg
accgcgcccg cctggagccg gccgagctgc tggccttcgt cgaagcgcgg 4080
gtgccgcgct acatgctccc ggcgcagttg cgcgtgctcg aacgcctgcc ggtcaccggc
4140 aacggcaaga tcgaccgcaa ggccctgacc ggctttgccc gccagcccca
ggcggacctt 4200 cggcatggcg tcgcgcaggc accggccgac gaactggaga
atgcgctgct ggcactctgg 4260 cgggaggtgc tggacaaccc gtcgctgggc
gtcgagcaag acttcttcgg ggccggcggc 4320 gactcgctgt tgatcgccca
gttgatcgcc cgtttgcgcg aacgactgga aagcgcccgt 4380 cggcatccgt
tcgatcgcct gctacgctgg gcgctcagcc agccgacgcc gcgcggcctg 4440
gccgaacgcc tgcgcagcgc gccggaagag ggccgtgggc cagccctggc cgcggcgcgc
4500 ggcgtcgccc cggcgccggc cggcatgtcg cgcgcaccgc tcgccgaggg
cgcggtggcg 4560 ctcgacccgc tggtgcgcct ggtgcccggc gagggcgtgc
cgcgggtgct ggtccacgaa 4620 ggcctcggca cgctactgcc gtaccgcccg
ctgcttcgcg ccctgggtga ggggcggccg 4680 ttgctggggc tggccgtgca
tgacagcgac gcctacctgg cgatccccgc cgagcatctc 4740 aacgcctgcc
tcggccgccg ctacgccgag gcgctccatc gcgccgggct acgcgaggtc 4800
gacctgctcg gctactgctc cggcgggctg gtcgccctgg agaccgccaa gtccctggtc
4860 cagcgcgggg tgcgcgtgcg ccaactggat atcgtctcca gctaccggat
tccctaccgg 4920 gtggacgacg agcgcctgct gttgttcagc ttcgccgcga
ccctcggcct ggataccgcg 4980 gcgctcggct tccccgcgcc ggaacgtctc
ggccaggcgg tgcaggcggc gctcgcgcag 5040 acaccggagc gcctggtcgc
cgaggcgctg gcggggctgc cgggcctggc cgatctcgtc 5100 gccctgcgcg
gccgcgtgct acaggcggcc agcggtagcg ccgacgccgt cagcgtcgaa 5160
cgcgacaccc tctaccggct gttctgtcac tcggtgcgtg ccagccaggc cgaggcgccg
5220 gagccctacg tcggcgcgct gcggctgttc gtgccggacg ccggcaaccc
attggtgccg 5280 cgctacgccg aggctctgga gacccaatgg cgggccgccg
cgcttggcgc gtgcggcatc 5340 cacgaggtgc ccggcgggca cttcgactgc
ctgggcgaag ccctggcgca atccttgtcg 5400 aaacccatgc cagaggaggc
gagccgatga 5430 32 1809 PRT Pseudomonas aeruginosa 32 Met Ser Leu
Gly Glu Leu Leu Glu Thr Cys Arg Ser Arg Arg Ile Glu 1 5 10 15 Leu
Trp Ser Glu Ala Gly Arg Leu Arg Tyr Arg Ala Pro Gln Gly Ala 20 25
30 Leu Asp Ala Gly Leu Ala Glu Arg Leu Arg Ala Glu Arg Glu Ala Leu
35 40 45 Leu Glu His Leu Glu Gly Gly Pro Gly Trp Arg Ala Glu Pro
Asp Met 50 55 60 Ala His Gln Arg Phe Pro Leu Thr Pro Val Gln Ala
Ala Tyr Val Leu 65 70 75 80 Gly Arg Gln Ala Ala Phe Asp Tyr Gly Gly
Asn Ala Cys Gln Leu Tyr 85 90 95 Ala Glu Tyr Asp Trp Pro Ala Asp
Thr Asp Pro Ala Arg Leu Glu Ala 100 105 110 Ala Trp Asn Ala Met Val
Glu Arg His Pro Met Leu Arg Ala Val Ile 115 120 125 Glu Asp Asn Ala
Trp Gln Arg Val Leu Pro Glu Val Pro Trp Gln Arg 130 135 140 Leu Thr
Val His Ala Cys Ala Gly Leu Asp Glu Ala Ala Phe Gln Ala 145 150 155
160 His Leu Glu Arg Val Arg Glu Arg Leu Asp His Ala Cys Ala Ala Leu
165 170 175 Asp Gln Trp Pro Val Leu Arg Pro Glu Leu Ser Ile Gly Arg
Asp Ala 180 185 190 Cys Val Leu His Cys Ser Val Asp Phe Thr Leu Val
Asp Tyr Ala Ser 195 200 205 Leu Gln Leu Leu Leu Gly Glu Trp Arg Arg
Arg Tyr Leu Asp Pro Gln 210 215 220 Trp Thr Ala Glu Pro Leu Glu Ala
Thr Phe Arg Asp Tyr Val Gly Val 225 230 235 240 Glu Gln Arg Arg Arg
Gln Ser Pro Ala Trp Gln Arg Asp Arg Asp Trp 245 250 255 Trp Leu Ala
Arg Leu Asp Ala Leu Pro Gly Arg Pro Asp Leu Pro Leu 260 265 270 Arg
Val Gln Pro Asp Thr Arg Ser Thr Arg Phe Arg His Phe His Ala 275 280
285 Arg Leu Asp Glu Ala Ala Trp Gln Ala Leu Gly Ala Arg Ala Gly Glu
290 295 300 His Gly Leu Ser Ala Ala Gly Val Ala Leu Ala Ala Phe Ala
Glu Thr 305 310 315 320 Ile Gly Arg Trp Ser Gln Ala Pro Ala Phe Cys
Leu Asn Leu Thr Val 325 330 335 Leu Asn Arg Pro Pro Leu His Pro Gln
Leu Ala Gln Val Leu Gly Asp 340 345 350 Phe Thr Ala Leu Ser Leu Leu
Ala Val Asp Ser Arg His Gly Asp Ser 355 360 365 Phe Val Glu Arg Ala
Arg Arg Ile Gly Glu Gln Met Phe Asp Asp Leu 370 375 380 Asp His Pro
Thr Phe Ser Gly Val Asp Leu Leu Arg Glu Leu Ala Arg 385 390 395 400
Arg Arg Gly Arg Gly Ala Asp Leu Met Pro Val Val Phe Thr Ser Gly 405
410 415 Ile Gly Ser Val Gln Arg Leu Leu Gly Asp Gly Glu Ala Pro Arg
Ala 420 425 430 Pro Arg Tyr Met Ile Ser Gln Thr Pro Gln Val Trp Leu
Asp Cys Gln 435 440 445 Val Thr Asp Gln Phe Gly Gly Leu Glu Ile Gly
Trp Asp Val Arg Leu 450 455 460 Gly Leu Phe Pro Glu Gly Gln Ala Glu
Ala Met Phe Asp Asp Phe Val 465 470 475 480 Gly Leu Leu Arg Arg Leu
Ala Gln Ser Pro Arg Ala Trp Thr Asp Gly 485 490 495 Asp Ala Thr Glu
Pro Val Glu Ala Pro Pro Gln Ala Leu Pro Gly Ser 500 505 510 Ala Arg
Ser Ile Ala Ala Gly Phe Ala Glu Arg Ala Leu Leu Thr Pro 515 520 525
Asp Ala Thr Ala Ile His Asp Ala Ala Gly Ser Tyr Ser Tyr Arg Gln 530
535 540 Val Ala Gln His Ala Ser Ala Leu Arg Arg Val Leu Glu Ala His
Gly 545 550 555 560 Ala Gly Arg Gly Arg Arg Val Ala Val Met Leu Pro
Lys Ser Ala Ala 565 570 575 Gln Leu Val Ala Val Ile Gly Ile Leu Gln
Ala Gly Ala Ala Tyr Val 580 585 590 Pro Val Asp Ile Arg Gln Pro Pro
Leu Arg Arg Gln Ala Ile Leu Ala 595 600 605 Ser Ala Glu Val Val Ala
Leu Val Cys Leu Glu Ser Asp Val Pro Asp 610 615 620 Val Gly Cys Ala
Cys Val Ala Ile Asp Arg Leu Ala Ala Asp Ser Ala 625 630 635 640 Trp
Pro Pro Pro Pro Ala Ala Glu Val Ala Ala Asp Asp Leu Ala Tyr 645 650
655 Val Ile Tyr Thr Ser Gly Ser Thr Gly Thr Pro Lys Gly Val Met Leu
660 665 670 Ser His Ala Ala Val Ser Asn Thr Leu Leu Asp Ile Asn Gln
Arg Tyr 675 680 685 Gly Val Asp Ala Asn Asp Arg Val Leu Gly Leu Ala
Glu Leu Ser Phe 690 695 700 Asp Leu Ser Val Tyr Asp Phe Phe Gly Ala
Thr Ala Ala Gly Ala Gln 705 710 715 720 Val Val Leu Pro Asp Pro Ala
Arg Gly Ser Asp Pro Ser His Trp Ala 725 730 735 Glu Leu Leu Glu Arg
His Ala Ile Thr Leu Trp Asn Ser Val Pro Ala 740 745 750 Gln Gly Gln
Met Leu Ile Asp Tyr Leu Glu Ser Glu Pro Gln Arg His 755 760 765 Leu
Pro Gly Pro Arg Cys Val Leu Trp Ser Gly Asp Trp Ile Pro Val 770 775
780 Ser Leu Pro Thr Arg Trp Trp Arg Arg Trp Pro Asp Ser Ala Leu Phe
785 790 795 800 Ser Leu Gly Gly Ala Thr Glu Ala Ala Ile Trp Ser Ile
Glu Gln Pro 805 810 815 Ile Arg Pro Gln His Thr Glu Leu Ala Ser Ile
Pro Tyr Gly Arg Ala 820 825 830 Leu Arg Gly Gln Ser Val Glu Val Leu
Asp Ala Arg Gly Arg Arg Cys 835 840 845 Pro Pro Gly Val Arg Gly Glu
Ile His Ile Gly Gly Val Gly Leu Ala 850 855 860 Leu Gly Tyr Ala Gly
Asp Pro Gln Arg Thr Ala Glu Arg Phe Val Arg 865 870 875 880 His Pro
Asp Gly Arg Arg Leu Tyr Arg Thr Gly Asp Leu Gly Arg Tyr 885 890 895
Leu Ala Asp Gly Ser Ile Glu Phe Leu Gly Arg Glu Asp Asp Gln Val 900
905 910 Lys Ile Arg Gly His Arg Ile Glu Leu Ala Glu Leu Asp Ala Ala
Leu 915 920 925 Cys Ala His Pro Gln Val Asn Leu Ala Ala Thr Val Val
Leu Gly Glu 930 935 940 Thr His Glu Arg Ser Leu Ala Ser Phe Val Thr
Leu His Ala Pro Val 945 950 955 960 Glu Ala Gly Glu Asp Pro Arg Thr
Ala Leu Asp Ala Val Arg Gln Arg 965 970 975 Ala Ala Gln Ala Leu Arg
Arg Asp Trp Gly Ser Glu Glu Gly Ile Ala 980 985 990 Ala Ala Val Ala
Ala Leu Asp Arg Ala Cys Leu Ala Ser Leu Ala Ala 995 1000 1005 Trp
Leu Ala Gly Ser Gly Leu Phe Ala Ser Ala Thr Pro Leu Asp 1010 1015
1020 Leu Ala Thr Leu Cys Gln Arg Leu Gly Ile Ala Glu Ala Arg Gln
1025 1030 1035 Arg Leu Leu Arg His Trp Leu Arg Gln Leu Glu Glu Gly
Gly Tyr 1040 1045 1050 Leu Arg Ala Glu Gly Glu Gly Trp Leu Gly Cys
Ala Glu Arg Pro 1055 1060 1065 Ala Gln Ser Pro Glu Asp Ala Trp Thr
Ala Phe Ala Gly Cys Ala 1070 1075 1080 Pro Ala Ala Leu Trp Pro Ala
Glu Leu Val Ala Tyr Leu Arg Asp 1085 1090 1095 Ser Ala Gln Ser Leu
Gly Glu Gln Leu Ala Gly Arg Ile Ser Pro 1100 1105 1110 Ala Ala Leu
Met Phe Pro Gln Gly Ser Ala Arg Ile Ala Glu Ala 1115 1120 1125 Met
Tyr Ser Gln Gly Leu His Ala Gln Ala Leu His Glu Ala Met 1130 1135
1140 Ala Glu Ala Ile Ala Ala Ile Val Glu Arg Gln Pro Gln Arg Arg
1145 1150 1155 Trp Arg Leu Leu Glu Leu Gly Ala Gly Thr Ala Ala Ala
Ser Arg 1160 1165 1170 Thr Val Ile Ala Arg Leu Ala Pro Leu Val Gln
Arg Gly Ala Glu 1175 1180 1185 Val Asp Tyr Leu Phe Thr Asp Val Ser
Ser Tyr Phe Leu Ala Ala 1190 1195 1200 Ala Arg Glu Arg Phe Ala Asp
Gln Pro Trp Val Arg Phe Gly Arg 1205 1210 1215 Phe Asp Met Asn Gly
Asp Leu Leu Asp Gln Gly Val Ala Pro His 1220 1225 1230 Ser Val Asp
Ile Leu Leu Ser Ser Gly Ala Leu Asn Asn Ala Leu 1235 1240 1245 Asp
Thr Pro Ala Leu Leu Ala Gly Leu Arg Glu Leu Leu Ser Ala 1250 1255
1260 Asp Ala Trp Leu Val Ile Gln Glu Leu Thr Arg Glu His Asn Glu
1265 1270 1275 Ile Ser Val Ser Gln Ser Leu Met Met Glu Asn Pro Arg
Asp Leu 1280 1285 1290 Arg Asp Glu Arg Arg Gln Leu Phe Val His Thr
Gly Gln Trp Leu 1295 1300 1305 Glu Trp Leu Ala Ala Gln Gly Gly Asp
Leu Ala Cys Gly Val Val 1310 1315 1320 Pro Pro Gly Ser Ala Leu Asp
Leu Leu Gly Tyr Asp Val Leu Leu 1325 1330 1335 Ala Arg Cys Lys Thr
Asp Arg Ala Arg Leu Glu Pro Ala Glu Leu 1340 1345 1350 Leu Ala Phe
Val Glu Ala Arg Val Pro Arg Tyr Met Leu Pro Ala 1355 1360 1365 Gln
Leu Arg Val Leu Glu Arg Leu Pro Val Thr Gly Asn Gly Lys 1370 1375
1380 Ile Asp Arg Lys Ala Leu Thr Gly Phe Ala Arg Gln Pro Gln Ala
1385 1390 1395 Asp Leu Arg His Gly Val Ala Gln Ala Pro Ala Asp Glu
Leu Glu 1400 1405 1410 Asn Ala Leu Leu Ala Leu Trp Arg Glu Val Leu
Asp Asn Pro Ser 1415 1420 1425 Leu Gly Val Glu Gln Asp Phe Phe Gly
Ala Gly Gly Asp Ser Leu 1430 1435 1440 Leu Ile Ala Gln Leu Ile Ala
Arg Leu Arg Glu Arg Leu Glu Ser 1445 1450 1455 Ala Arg Arg His Pro
Phe Asp Arg Leu Leu Arg Trp Ala Leu Ser 1460 1465 1470 Gln Pro Thr
Pro Arg Gly Leu Ala Glu Arg Leu Arg Ser Ala Pro 1475 1480 1485 Glu
Glu Gly Arg Gly Pro Ala Leu Ala Ala Ala Arg Gly Val Ala 1490 1495
1500 Pro Ala Pro Ala Gly Met Ser Arg Ala Pro Leu Ala Glu Gly Ala
1505 1510 1515 Val Ala Leu Asp Pro Leu Val Arg Leu Val Pro Gly Glu
Gly Val 1520 1525 1530 Pro Arg Val Leu Val His Glu Gly Leu Gly Thr
Leu Leu Pro Tyr 1535 1540 1545 Arg Pro Leu Leu Arg Ala Leu Gly Glu
Gly Arg Pro Leu Leu Gly 1550 1555 1560 Leu Ala Val His Asp Ser Asp
Ala Tyr Leu Ala Ile Pro Ala Glu 1565 1570 1575 His Leu Asn Ala Cys
Leu Gly Arg Arg Tyr Ala Glu Ala Leu His 1580 1585 1590 Arg Ala Gly
Leu Arg Glu Val Asp Leu Leu Gly Tyr Cys Ser Gly 1595
1600 1605 Gly Leu Val Ala Leu Glu Thr Ala Lys Ser Leu Val Gln Arg
Gly 1610 1615 1620 Val Arg Val Arg Gln Leu Asp Ile Val Ser Ser Tyr
Arg Ile Pro 1625 1630 1635 Tyr Arg Val Asp Asp Glu Arg Leu Leu Leu
Phe Ser Phe Ala Ala 1640 1645 1650 Thr Leu Gly Leu Asp Thr Ala Ala
Leu Gly Phe Pro Ala Pro Glu 1655 1660 1665 Arg Leu Gly Gln Ala Val
Gln Ala Ala Leu Ala Gln Thr Pro Glu 1670 1675 1680 Arg Leu Val Ala
Glu Ala Leu Ala Gly Leu Pro Gly Leu Ala Asp 1685 1690 1695 Leu Val
Ala Leu Arg Gly Arg Val Leu Gln Ala Ala Ser Gly Ser 1700 1705 1710
Ala Asp Ala Val Ser Val Glu Arg Asp Thr Leu Tyr Arg Leu Phe 1715
1720 1725 Cys His Ser Val Arg Ala Ser Gln Ala Glu Ala Pro Glu Pro
Tyr 1730 1735 1740 Val Gly Ala Leu Arg Leu Phe Val Pro Asp Ala Gly
Asn Pro Leu 1745 1750 1755 Val Pro Arg Tyr Ala Glu Ala Leu Glu Thr
Gln Trp Arg Ala Ala 1760 1765 1770 Ala Leu Gly Ala Cys Gly Ile His
Glu Val Pro Gly Gly His Phe 1775 1780 1785 Asp Cys Leu Gly Glu Ala
Leu Ala Gln Ser Leu Ser Lys Pro Met 1790 1795 1800 Pro Glu Glu Ala
Ser Arg 1805 33 1713 DNA Pseudomonas aeruginosa 33 gtgaccccgg
tgctgtggcg cctgctgcgc acctatcgct ggcggctggc ggcggccatg 60
gggttgcagg ccctggccgg gctctgctcg ctgttgccct ggatgcttct cgcctggctc
120 gccgagccgc tggcgcgcgg ccaggcgcag ccggccctgc tggccctggt
gctgctggcg 180 gtgctggcct ggctgggctg ccaggcgctg gccgcgcacc
tggcccaccg ggtcgacgcg 240 gacctctgca acgacctgcg cctgcgcctg
ctggcgcacc tgcaacggct gccgctggac 300 tggttcggtc gccagggccc
ggacggcgtg gcgcgcctcg tggagcagga cgtgcgggcc 360 ctgcaccaac
tgatcgcgca cgctcccaac gatctcagca acctgttggt ggtgccgctc 420
gtcgcgttgc tctggctggc ctggctgcac ccctggctgc tgctgttctg cctgctgccg
480 ctggtgctgg ccgccgccgg cttcctgctg ctgcgctcgg cgcgctaccg
cgacctggtg 540 ctgcggcgca acgccgcgct ggaaaggctc tcggcggact
atggcgaatt cgcccacaac 600 ctgctgctgg cccgacagta ccccggcgcc
ggcatacaac agggcgccga ggcgtcggcg 660 gcggccttcg gcgaagcgtt
cggcgcctgg gtgaagcggg tcggccacct cgccgcgctg 720 gtctacgtgc
agttgtcgac gccctggctg ctggcctggg tcctgctcgg cgcgctggcc 780
ctggatgccc tcggcgtgcc gctggcgctc ggccaggcct gtgccttcct gctcctgctg
840 cgggccttgg ctgccccggt gcaggcgctc ggccacggcg gcgacgcgct
gctgggcgcg 900 cgcgccgccg ccgagcgcct gcagcaggtg ttcgaccagg
cgccgctggc cgagggccgc 960 tcgacccgcg agccggtcga tggcgcggtg
gcgctgcacg gcctgggcca tgcctatgaa 1020 ggcgtggagg tcctggccga
tatcgatctg gagctggagg atggcagcct ggtggccctg 1080 gtcggtccct
cgggctccgg caagagcacc ctgctgcacc tgctggcgcg ctacatggac 1140
gcgcagcgcg gcgaactgga ggttggcggc ctggcactga aggacatgcc tgatgccgtg
1200 cgccatcggc atatcgcgct ggtcggccag caggcggcgg cgctggagat
atccctggcc 1260 gacaacattg ccctgttccg ccccgatgcc gatctccagg
agattcgcca ggcggcccgt 1320 gacgcctgcc tcgacgagcg catcatggcc
ctgccgcgtg gctacgacag cgtgccggga 1380 cgcgacctgc aactgtccgg
cggcgaactg caacgactgg ccctggcccg tgcgctgcta 1440 tcgccggcga
gcctgttgct gctcgacgag ccaacctcgg cgctggatcc gcagaccgcc 1500
cggcaggtcc tgcgcaacct gcgcgaacgc ggcggtggcc ggacccgggt gatcgtcgcc
1560 catcgtctgg ccgaagtcag cgatgccgac ctgatcctgg tgctggtcgc
tggccgtctg 1620 gtcgaacgcg gcgagcacgc ggcgctgttg gcggcggacg
gcgcctatgc gcgcttgtgg 1680 cgtgaacaga acggcgcgga ggtggcggca tga
1713 34 570 PRT Pseudomonas aeruginosa 34 Met Thr Pro Val Leu Trp
Arg Leu Leu Arg Thr Tyr Arg Trp Arg Leu 1 5 10 15 Ala Ala Ala Met
Gly Leu Gln Ala Leu Ala Gly Leu Cys Ser Leu Leu 20 25 30 Pro Trp
Met Leu Leu Ala Trp Leu Ala Glu Pro Leu Ala Arg Gly Gln 35 40 45
Ala Gln Pro Ala Leu Leu Ala Leu Val Leu Leu Ala Val Leu Ala Trp 50
55 60 Leu Gly Cys Gln Ala Leu Ala Ala His Leu Ala His Arg Val Asp
Ala 65 70 75 80 Asp Leu Cys Asn Asp Leu Arg Leu Arg Leu Leu Ala His
Leu Gln Arg 85 90 95 Leu Pro Leu Asp Trp Phe Gly Arg Gln Gly Pro
Asp Gly Val Ala Arg 100 105 110 Leu Val Glu Gln Asp Val Arg Ala Leu
His Gln Leu Ile Ala His Ala 115 120 125 Pro Asn Asp Leu Ser Asn Leu
Leu Val Val Pro Leu Val Ala Leu Leu 130 135 140 Trp Leu Ala Trp Leu
His Pro Trp Leu Leu Leu Phe Cys Leu Leu Pro 145 150 155 160 Leu Val
Leu Ala Ala Ala Gly Phe Leu Leu Leu Arg Ser Ala Arg Tyr 165 170 175
Arg Asp Leu Val Leu Arg Arg Asn Ala Ala Leu Glu Arg Leu Ser Ala 180
185 190 Asp Tyr Gly Glu Phe Ala His Asn Leu Leu Leu Ala Arg Gln Tyr
Pro 195 200 205 Gly Ala Gly Ile Gln Gln Gly Ala Glu Ala Ser Ala Ala
Ala Phe Gly 210 215 220 Glu Ala Phe Gly Ala Trp Val Lys Arg Val Gly
His Leu Ala Ala Leu 225 230 235 240 Val Tyr Val Gln Leu Ser Thr Pro
Trp Leu Leu Ala Trp Val Leu Leu 245 250 255 Gly Ala Leu Ala Leu Asp
Ala Leu Gly Val Pro Leu Ala Leu Gly Gln 260 265 270 Ala Cys Ala Phe
Leu Leu Leu Leu Arg Ala Leu Ala Ala Pro Val Gln 275 280 285 Ala Leu
Gly His Gly Gly Asp Ala Leu Leu Gly Ala Arg Ala Ala Ala 290 295 300
Glu Arg Leu Gln Gln Val Phe Asp Gln Ala Pro Leu Ala Glu Gly Arg 305
310 315 320 Ser Thr Arg Glu Pro Val Asp Gly Ala Val Ala Leu His Gly
Leu Gly 325 330 335 His Ala Tyr Glu Gly Val Glu Val Leu Ala Asp Ile
Asp Leu Glu Leu 340 345 350 Glu Asp Gly Ser Leu Val Ala Leu Val Gly
Pro Ser Gly Ser Gly Lys 355 360 365 Ser Thr Leu Leu His Leu Leu Ala
Arg Tyr Met Asp Ala Gln Arg Gly 370 375 380 Glu Leu Glu Val Gly Gly
Leu Ala Leu Lys Asp Met Pro Asp Ala Val 385 390 395 400 Arg His Arg
His Ile Ala Leu Val Gly Gln Gln Ala Ala Ala Leu Glu 405 410 415 Ile
Ser Leu Ala Asp Asn Ile Ala Leu Phe Arg Pro Asp Ala Asp Leu 420 425
430 Gln Glu Ile Arg Gln Ala Ala Arg Asp Ala Cys Leu Asp Glu Arg Ile
435 440 445 Met Ala Leu Pro Arg Gly Tyr Asp Ser Val Pro Gly Arg Asp
Leu Gln 450 455 460 Leu Ser Gly Gly Glu Leu Gln Arg Leu Ala Leu Ala
Arg Ala Leu Leu 465 470 475 480 Ser Pro Ala Ser Leu Leu Leu Leu Asp
Glu Pro Thr Ser Ala Leu Asp 485 490 495 Pro Gln Thr Ala Arg Gln Val
Leu Arg Asn Leu Arg Glu Arg Gly Gly 500 505 510 Gly Arg Thr Arg Val
Ile Val Ala His Arg Leu Ala Glu Val Ser Asp 515 520 525 Ala Asp Leu
Ile Leu Val Leu Val Ala Gly Arg Leu Val Glu Arg Gly 530 535 540 Glu
His Ala Ala Leu Leu Ala Ala Asp Gly Ala Tyr Ala Arg Leu Trp 545 550
555 560 Arg Glu Gln Asn Gly Ala Glu Val Ala Ala 565 570 35 1725 DNA
Pseudomonas aeruginosa 35 atgaccctgt tcgaacgaat gcgtgcgctg
cccgaagact gccgtgccgc gttgcgccgg 60 gcgagcgcct gggcggtcct
ggcggcgctg ctggacgccg cttgcggcgt attgctggtg 120 ccgttggtcg
aggcctggtt cgccgaaggc gcgttgccct ggcgctgggt cgccgcgttg 180
ctcggcttga gcctggcgca ggcgctgttg cagtacctgg ccctgcgtcg cggtttcgcc
240 gccggcggct cgctggcggc tggactggtg cgcagcctgg tggcgcgctt
gccgcgcctg 300 gcgccgccgg cgctgcgccg ggtcgcgccg gccgaaggcc
tgctgcgcgg cccggtgatg 360 caggcgatgg gcattccggc gcacctgctg
gggccgctga tcgccgcgtt ggtgacgccg 420 ctcggggtga tcctcgggct
gttcctgatc gacccgtcca tcgccctcgg cctgctcctt 480 gctggtgcct
tcctcgccgc gctgttgcgc tggagcgggc ggcgcaatct ggcggcggag 540
gatgcccggc tggccgccga gcgcgacgcc gcacggcagt tgcaggcgtt cgccgaacgc
600 cagccactgc tgcgcgcggc gcagcgcgaa agcgtcgccc gccaggggct
ggaagaggcc 660 ttgcgcagtc tccaccgcag caccctggat ctgttgcggc
gcagcctgcc cagcggcctc 720 ggcttcgccc tggcggtgca ggcggcgttc
gccttcgccc tgctcggcgg cgcctgggcg 780 gtggagcggc aatggctgga
cggcgctcgg ctggtggccg tgctggtgct gctggtgcgc 840 ttcatcgagc
cgctggccca gctcacccat ctcgaccagg cgttgcgcgg cgcctggcag 900
gcgctggata ccctgctgcg ggttttcgcc ctggctccgc tgcgcagccc cgagccgggc
960 gagcggccgc acgacgccag cctggcggcc gaggccgtgg aattgcgcct
ggaagatggc 1020 cgcgccttgc tcgaggacat ttccctgagg ctggagccgg
gttcgctgaa cgtcctcgtc 1080 ggaccctccg gggccggcaa gagcagcctg
ctggcgctgc tcgggcggct ctacgacgtc 1140 gatgccgggc gtgtcctgct
gggtggcgtg gatatccgcc ggttgagcga aacgaccctc 1200 gccgccagtc
gtaacctggt gttccaggac aacggcctgt tccgcggcag cgttgcctgg 1260
aacctgcgca tggcgcgagc ggacgccgat ctcgaagcgc tgcgcgaggc ggcgcgggcg
1320 gttggcctgc tggaagagat cgaggcctgg ccgcagggct gggacagcga
cgtcggtccc 1380 ggcggcgcgc tgctgtccgg cggccagcgg caacgcctgt
gcctggctcg cgggctgctc 1440 tcgacggcgc cgttgctgct gctcgacgag
cccaccgcca gcctcgacgc cgccagcgag 1500 gcgcaggtgc tgcgcagcct
gctcgggttg cgcggccggc gcaccctgct ggtagtgacc 1560 caccgcccgg
cgctggcgcg tcaggccgac caggtactgc tgctggagga ggggcgcctg 1620
cgcctcagcg gacttcacgc cgatctgctc gtccgggacg actggtatgc cggtttcgtc
1680 gggctggcgg gcgaggaaag ttccgcgacg gtcgtggatc gatag 1725 36 574
PRT Pseudomonas aeruginosa 36 Met Thr Leu Phe Glu Arg Met Arg Ala
Leu Pro Glu Asp Cys Arg Ala 1 5 10 15 Ala Leu Arg Arg Ala Ser Ala
Trp Ala Val Leu Ala Ala Leu Leu Asp 20 25 30 Ala Ala Cys Gly Val
Leu Leu Val Pro Leu Val Glu Ala Trp Phe Ala 35 40 45 Glu Gly Ala
Leu Pro Trp Arg Trp Val Ala Ala Leu Leu Gly Leu Ser 50 55 60 Leu
Ala Gln Ala Leu Leu Gln Tyr Leu Ala Leu Arg Arg Gly Phe Ala 65 70
75 80 Ala Gly Gly Ser Leu Ala Ala Gly Leu Val Arg Ser Leu Val Ala
Arg 85 90 95 Leu Pro Arg Leu Ala Pro Pro Ala Leu Arg Arg Val Ala
Pro Ala Glu 100 105 110 Gly Leu Leu Arg Gly Pro Val Met Gln Ala Met
Gly Ile Pro Ala His 115 120 125 Leu Leu Gly Pro Leu Ile Ala Ala Leu
Val Thr Pro Leu Gly Val Ile 130 135 140 Leu Gly Leu Phe Leu Ile Asp
Pro Ser Ile Ala Leu Gly Leu Leu Leu 145 150 155 160 Ala Gly Ala Phe
Leu Ala Ala Leu Leu Arg Trp Ser Gly Arg Arg Asn 165 170 175 Leu Ala
Ala Glu Asp Ala Arg Leu Ala Ala Glu Arg Asp Ala Ala Arg 180 185 190
Gln Leu Gln Ala Phe Ala Glu Arg Gln Pro Leu Leu Arg Ala Ala Gln 195
200 205 Arg Glu Ser Val Ala Arg Gln Gly Leu Glu Glu Ala Leu Arg Ser
Leu 210 215 220 His Arg Ser Thr Leu Asp Leu Leu Arg Arg Ser Leu Pro
Ser Gly Leu 225 230 235 240 Gly Phe Ala Leu Ala Val Gln Ala Ala Phe
Ala Phe Ala Leu Leu Gly 245 250 255 Gly Ala Trp Ala Val Glu Arg Gln
Trp Leu Asp Gly Ala Arg Leu Val 260 265 270 Ala Val Leu Val Leu Leu
Val Arg Phe Ile Glu Pro Leu Ala Gln Leu 275 280 285 Thr His Leu Asp
Gln Ala Leu Arg Gly Ala Trp Gln Ala Leu Asp Thr 290 295 300 Leu Leu
Arg Val Phe Ala Leu Ala Pro Leu Arg Ser Pro Glu Pro Gly 305 310 315
320 Glu Arg Pro His Asp Ala Ser Leu Ala Ala Glu Ala Val Glu Leu Arg
325 330 335 Leu Glu Asp Gly Arg Ala Leu Leu Glu Asp Ile Ser Leu Arg
Leu Glu 340 345 350 Pro Gly Ser Leu Asn Val Leu Val Gly Pro Ser Gly
Ala Gly Lys Ser 355 360 365 Ser Leu Leu Ala Leu Leu Gly Arg Leu Tyr
Asp Val Asp Ala Gly Arg 370 375 380 Val Leu Leu Gly Gly Val Asp Ile
Arg Arg Leu Ser Glu Thr Thr Leu 385 390 395 400 Ala Ala Ser Arg Asn
Leu Val Phe Gln Asp Asn Gly Leu Phe Arg Gly 405 410 415 Ser Val Ala
Trp Asn Leu Arg Met Ala Arg Ala Asp Ala Asp Leu Glu 420 425 430 Ala
Leu Arg Glu Ala Ala Arg Ala Val Gly Leu Leu Glu Glu Ile Glu 435 440
445 Ala Trp Pro Gln Gly Trp Asp Ser Asp Val Gly Pro Gly Gly Ala Leu
450 455 460 Leu Ser Gly Gly Gln Arg Gln Arg Leu Cys Leu Ala Arg Gly
Leu Leu 465 470 475 480 Ser Thr Ala Pro Leu Leu Leu Leu Asp Glu Pro
Thr Ala Ser Leu Asp 485 490 495 Ala Ala Ser Glu Ala Gln Val Leu Arg
Ser Leu Leu Gly Leu Arg Gly 500 505 510 Arg Arg Thr Leu Leu Val Val
Thr His Arg Pro Ala Leu Ala Arg Gln 515 520 525 Ala Asp Gln Val Leu
Leu Leu Glu Glu Gly Arg Leu Arg Leu Ser Gly 530 535 540 Leu His Ala
Asp Leu Leu Val Arg Asp Asp Trp Tyr Ala Gly Phe Val 545 550 555 560
Gly Leu Ala Gly Glu Glu Ser Ser Ala Thr Val Val Asp Arg 565 570 37
558 DNA Pseudomonas aeruginosa misc_feature (85)..(85) n is a, c,
g, or t 37 ctctttcagc cgcacgcggc gcacctcgtg tgtgatcagt gagtggtttg
caactgcggg 60 tcaaggatct ggatttccct cacangtncg atcatcgtgc
gggagggcaa gggctccaag 120 gatcgggcct tgatgttacc cgagagcttg
gcacccagcc tgcgcgagca gggnnaattg 180 atccggtgga tgaccttttg
aatgaccttt aatagattat attactaatt aattggggac 240 cctanaggtc
ccctttttta ttttaaaaat tttttcacaa aacggtttat ttncataaag 300
cttgctcaat caatcaccnt atccncggga attcggccta ggcggccaga tctgatcaag
360 agacagacct ccagctttgc atccggagcg accacacgag cgaggtcagt
cactttcatc 420 gaaggaattt tcttgacata gatctcacca ccttccatgt
cctcaaaggc atgccacact 480 aactcgacgc cctcctccaa agaaatcatg
aaccgggtca tccgctcatc agtgataggc 540 aagacgccct tgtccttg 558 38 479
DNA Klebsiella sp. 38 acgcaggata tcttcttcat caaattgtcg atgcccgcct
tcgctacgct gcggtttcag 60 tagaccgtaa cgacgctgcc aggcgcgcag
tgtgaccgga ttgattccgc aacgttcggc 120 gacttcaccg atactgtaaa
acgccatagc agcctcacat caacctgata ccttaatacc 180 taaactaacg
aattcaggca tcctgtacaa ctctattttc ttgtacagat aaagatatca 240
ggttgcggct cacagcgccc gggaaaaaag atgaaaaaat gtttagctga tttcgcggtg
300 gttcattttt tctccggcca tgcgacggcg ggtaggcccc ccaggcgcgc
gctggcgaac 360 aaattgccct gaaactgtga aataccggct gattccagcc
acatccactc ttcagcacgc 420 tcaacgccga cggctgagac cgcaatctcc
agagaagtac agcatttgat aatcgcctg 479 39 516 DNA Klebsiella sp. 39
gaccatgtgc tgatgaccaa taccgcctat gagccaagcc aggacttttg taccaaaatt
60 ctcgccaaac tcggcgtcac caccagctgg ttcgatccct taatcggcgc
cgatatcgcc 120 cgtctggttc gccctgagac ccgcgtggtg ttcctcgaat
cgcccggctc gatcaccatg 180 gaagtgcacg atgtgccggc gatagtcgcc
gccgtgcgtc aggtcgcccc ggaagcgatt 240 atcatgatcg ataacacctg
ggcggcgggg atcctgttta aagccctgga ttttggcatt 300 gatatttcca
ttcaggcagg caccaaatac ctgatcggcc attccgacgc catggtgggc 360
accgcggtgg cgaacgcgcg ctgctggccg cagctgcgtg aaaatgccta cctgatgggg
420 caaatgctgg acgccgatac tgcctatatg accagccgcg gcctgcgaac
cctgggcgtg 480 cgcctgcgtc agcatcatga aagcagcctg cgcatc 516 40 377
DNA Klebsiella sp. 40 cttttggccc cttttttgtc tttattctgg agaacttatt
atggcgaaag aatttggtcg 60 cccgcagcgt gtggcccagg agatgcaaaa
agagattgcc atcatcctgc agcgtgaaat 120 taaagatccg cgtctgggca
tgatgaccac cgtttccggt gtggaaatgt cccgtgacct 180 ggcctatgcc
aaggtgtatg tcaccttcct taacgacaaa gatgaagccg cggtgaaagc 240
gggcatcaaa gcgctgcagg aagcttctgg ctttatccgc tctctgctgg ggaaagcgat
300 gcgtctgcgc atcgtaccgg aactgacttt cttctacgac aactcactgg
tggaagggat 360 gcgtatgtcc aacctgg 377 41 625 DNA Klebsiella sp. 41
gcccagcccg ctttcccgct tgcccagtta aaagccttcg tggagcagga atttgctcag
60 attaagcatg ttctgcacgg catcagcctg ctgggtcagt gcccggacag
cgtcaatgcc 120 gcgctgatct gccgcggcga aaagctctcc atcgccatca
tggcgggtct gctggaagcc 180 cgtggacaca aagtcagtgt cattaacccg
gtcgaaaaac tgctcgccgt gggtcactat 240 ctggaatcca ccgtcgatat
cgccgaatcc acccgccgca ttgccgccag ccagatcccg 300 gcagaccata
tgatcctgat ggccgggttt accgccggca atgagaaagg cgagctggtg 360
gtgctggggc gtaacggctc cgactactcg gctgcggtac tggccgcctg cctgcgcgct
420 gactgctgcg aaatctggac cgatgtcgac ggagtgtaca cctgcgatcc
gcgtcaggtg 480 ccggatgcgc gcctgctgaa atcgatgtct tatcaggagg
cgatggagct ctcctacttt 540 ggcgcgaaag tgctgcaccc gcgcaccatt
gcccctatcg cccagttcca aatcccatgc 600 ctgattaaaa ataccggcaa ccccc
625 42 355 DNA Klebsiella sp. 42 ggcgcagcgt ctgctcgtca ccgtcaagct
cgaagcttaa cattgcgcca aaaccttttt 60 gctgacgcgc cgcaatttca
tgcccctggt tttccggcag cgatggatga tacagctttt 120 tcaccagcgg
ctgggttttc agatactcaa cgatcgccag ggcatttcgc tgcgccactt 180
ccatccgtgg agacagcgtc cgcagcccgc gcaacagcag atagctgtcg aaggcgctgc
240 cggtgacgcc
aatattattc gcccaccatg ccagttcggt gacagttgcc ggatctttgg 300
caatcaccac cccggccacc acatcggagt gaccattgag gtatttggta cagga 355 43
500 DNA Klebsiella sp. 43 gttgcgtccc aggcgggtaa acgcatcctg
caggtagtca atttcgtcgt cggccagcgc 60 cagacccaga cggaggttgg
cgtcaatcag cgcctgacgc ccttcgccca gcaggtcgac 120 gctggtgacc
ggcgtcggct gatggtgagc gaacagcttc tcgcccgctt ccagctcgtc 180
gaagacgctc tccatcatgc ggtcatgcag ctccgccgcc accgcggccc actgcgcttc
240 ggtcagggtt gaggcttcaa cgtaatacgc cacgccgcgc tcaagacgca
caacctgcgc 300 cagaccgcag ttgtgagcga tatcggtagc tttagaagac
cagggagaga tggtgccagg 360 gcgaggggtc acgagcagta atttaccggt
cggggtatgg ctgcttaagc tcgggccata 420 ctgaagcagt cgcgccaggc
gctcgcgatc gtcagcgctc agcggggcgt tcagatcggc 480 aaaatgaata
tattcggcat 500 44 439 DNA Klebsiella sp. 44 gtattggcat cgtactcctg
ggctggccgg tgacaaaggc gatgcgctta tctttgctgg 60 cgaacaaata
cgcatcgccc tcttccgtct ccgcgaggat ctcgagatcg gtatagtcgc 120
gaataagtcc ggccggaaaa tcagcatagc gtgagtgcgg ggccaggaaa gagtcgtcga
180 aaccgcgggt cagtaaggcg tgcggatgaa gaatatggtg ttcatagacg
ccggaaatct 240 tttcggcgcg ggtctgcttg ggaatgccgt acagaatgtt
cagcgcggcc tgaaccgccc 300 aacagacgaa cagcgtcgaa gtgacgtgat
ccttggccca ctccagcacc tgtttgatct 360 gcggccagta agcaacatcg
ttaaactcaa ccaggcctaa aggagcgccg gtaacaatca 420 ggccgtcaaa
gttctgatc 439 45 297 DNA Klebsiella sp. 45 gaggttcata tgtccgtact
cgatctaaac gcgcttaatg cattgccgaa agtggaacgc 60 attctggcac
tcgcggaaac caacgcccaa ctggaaaagc ttgacgccga agggcgtgtg 120
gcgtgggcgc tggaaaatct gccgggaaac tatgtgctgt cgtcgagctt tggcattcag
180 gcggcggtaa gtttgcatct ggtgaatcag atccgcccgg acattccggt
gatcctcacc 240 gataccggct acctgttccc ggaaacctat cagtttattg
acgagctgac ggacaag 297 46 502 DNA Klebsiella sp. 46 tgttaaagcg
tgcgttctac agcctgttag tcctgctcgg cctgctgctg ttgaccgtgc 60
tgggccttga ccgctggatg agctggaaaa ccgcgcccta tatctatgat gaactgcagg
120 acctgcccta ccgtcaggtc ggtgtggtgc tgggcaccgc caaatattac
cgcaccggcg 180 tcatcaatca gtattaccgt taccgcatcc agggtgcgct
gaacgcctac aacagcggca 240 aggtcaacta tctcctgctg agcggcgata
atgctctgca aagctacaat gaaccgatga 300 ccatgcgtcg ggacctgatt
aaaggcggcg tcgatcccgc ggatatcgta ctggactatg 360 ccggtttccg
taccctcgac tcgatcgtcc gtacccggaa agtgttcgac accaacgact 420
tcattatcat cacccagcgc ttccactgcg aacgggcgct gtttatcgcc ctgcatatgg
480 ggatccaggc ccagtgctac gc 502 47 500 DNA Klebsiella sp. 47
cgctgaacct cctcaaacaa acgcaggccc tgcacctgtc ggctgcaggc gaccagcgtg
60 gatccgctca aacagctgca ggccgagcac cttctcaaag cgcgccagct
cgcggctgac 120 cgtgggttgc gaggtgtgca gcatccgcgc cgcttcggtc
aggttgccgg tggtcatcac 180 cgcgtgaaag atttcgatat gacgcaaatt
gacggctggc atgcggtctc cgtgaggctc 240 ggctggaacc atatcatttt
tgcatagagt cgcgataaaa cgatattttt tattcgtctg 300 tcactgtggc
gtaatcagaa aaaacagcga ccaacacacg cactgcaccg gagttcttat 360
gccacactcg ctttacgcca ccgatactga cctgaccgcg gacaacctgc tgcgcctgcc
420 ggcggaattt ggctgcccgg tctgggtcta tgatgcgcag attattcgcc
gccagatagc 480 ccagctcagc cagtttcgac 500 48 229 DNA Klebsiella sp.
48 ggcttccacc caaatcgctt tgtcggcaac gatttttgct aaaacggctt
tgcattcttt 60 accctcttgc ccgctaagtg cggtcactct gtcataggcc
gcgccgctgc tgcagcacat 120 ccagtacctg ctgagcgtta gctttcagat
cttcatgccc gtgtaaacgc atcaatatgg 180 cgacgttggc ggcgacggcg
gcttcgtgag cggcttcacc tttaccttg 229 49 466 DNA Klebsiella sp. 49
tggctcaacg ctgctcagtg gtgcgaggtg tcactttggt gatcacatcg gcgttgtctg
60 cacagtgaaa tcagatccag cgccgcgtcc ggttttacgc acgtagtccg
gattgtgggt 120 gcctttctta acgatattca gccacggccc ttcgagatgc
aggcccagcg cctggttcgg 180 atgtttttgc agatattcgc gcatcacgcg
cacgccttgc ttcatcagat cgtcgctgga 240 ggtaatcagc gtcggcagga
agctggtgca gcctgagcgt tcgttggcct tctgcatgat 300 ctccagcgtt
tcgacagtga ccgcctctgg gctgtcgtta aactgcacgc cgccgcagcc 360
gttgagctgg acgtcgataa aaccgggggc gattattgcg ccgttgactg agcgctgctc
420 gatgtcagac ggcaaatctg ccagcggaca aagacgttcg ataaag 466 50 450
DNA Klebsiella sp. 50 ttaagcacca tatcgtaccg ctgctggcgc agcgtctgaa
tgagctgcca ttgcatcttc 60 agctgatacc tttttccctg gctttttcca
gcggcgatcg agaccataaa tatggtggat 120 atcggggttg gctgcgagca
tatcccgggt ctcttcatac aacaggacat ccacgctggc 180 ggcggggtac
tgctgtttca gcgcgtgaat aagcggcgtg atcagcagca tgtcgccatg 240
atggcgcagc ttaatgacca ggatccgcgc cgggttcaac gggccgcggg agagggtttc
300 aggcgtcata ctctgttctt catccaggat aagggttccg attctagggg
atcagacaga 360 ttgagagaag cgttgtattg ctctaccatg acccgatacg
tatggcctga ggacgttttc 420 gtgcacaatc ccgcaatttc tcatcacgat 450 51
450 DNA Klebsiella sp. 51 cactcaggct tgcctgtaac gcttgttcgc
catcacgtaa ggtcgtatcg aaaataatga 60 cttgctggct catggtttgg
atccttagtc tgtgtcctgg cgccttgttg acgagcataa 120 aaaaacccgc
gccaaggcgc gggttttata gtcttgctgg aagatgactt aacgctgaac 180
gtcgcccaac agcctaccga gcaaatggca tgcgtttagt agtagtaggc tggtgatacg
240 agcggtgcga atcattgcgt caaactccag atgaaatcgt tatgctttta
gagttactgg 300 atagccgttt taaagtcaac ccctggcatg gaaaaagcgt
tttgggctga ctaaatgaat 360 tagcaaaatg tgctgatgta agccccattt
tgccgaagat cctattttgg accgaaggcg 420 gtttatcccc aatttgtttc
atttgaaaaa 450 52 575 DNA Klebsiella sp. 52 cgctgaaccg ctatccggag
ccgcagccga agtgccgtga ttgagagcta cgcccgctac 60 gccgaggtca
aaccggagca ggtgctggtc agccgcggcg ccgacgaagg catcgagctg 120
ctgatccgcg ccttctgtga gcccggcgaa gacgcggtgc tctactgccc gccgacctac
180 ggcatgtaca gcgtcagcgc cgagaccatc ggcgtcgagt gccgcaccgt
gccgacgctg 240 gccagctggc agctcgacct gccgggcatc gaagcgcggc
tggacggcgt gaaggtggtg 300 tttgtctgca gcccgaacaa cccgaccggg
cagattatcg acccgcagtc gatgcgcgac 360 ctgctggaga tgacccgcgg
caaagccatc gtggtggccg acgaagccta tattgaattc 420 tgcccgcagg
cgacgctcgc cggctggctc agcgactatc cgcacctggt ggtgctgcgc 480
acgctgtcca aagccttcgc cctcgccggc ctgcgctgcg gcttcaccct cgccaacgcc
540 gaggtgatta acgtgctgct gaaagtgatc gcccc 575 53 375 DNA
Klebsiella sp. 53 cgtatatttc atcgtacaga aaccgtaaac acaggcattg
gctgattttc agtgagtgaa 60 tttaaataga cttctgccgt tttcaatgct
tcggcgatgg tcacatccat atcaaggtaa 120 cggtaggttc caagacgacc
gacaaaagtg atgttggttt cattctcggc caatgacaaa 180 tatttttcaa
gaagagccat ttctcccatc tggcgaatag gatagtaagg aatatcattt 240
tcttcacaag cacggctata ctctttataa caaacagagc cgtcgtgttg ttcccaggga
300 gaaaaatatt tatgttcagt gatgcgagta tagggcacat ccacagaaca
gtagttcatc 360 actgcgcatc cctgg 375 54 400 DNA Klebsiella sp. 54
gtacgccgat tttatatgcg tctgatatga ttccggaaaa atttagctgg ataattacct
60 acaatccgct agcgagtatg attcttagtt ggcgtgattt attcatgaat
gggactctta 120 attttgagta tatttctata ctctatttta cgggaattat
tttgacggtt gtcggtttgt 180 ctattttcaa taaattaaaa tatcgatttg
cagagatcta aaagtgcgct ataagagcag 240 catgctaggc tatttatggt
cagtagcaaa tccattgctt tttgccatga tttactattt 300 tatatttaag
ctggtaatga gagtacaaat tccaaattat acagttttcc tcattaccgg 360
cttgtttccg tggcaatggt ttgccagttc ggccactaac 400 55 413 DNA
Klebsiella sp. 55 cgagccaccc actgtagcgt atggatatcg cgcaagccgc
cggggctgct tttcacgtcc 60 ggctcgaggt tatagctggt gccatgatag
cgctgatgac ggacgttctg ctcttcgacc 120 ttggcggcga agaacttttc
cgatggccag aagccgtcgc taaaaatatg tttttgcagt 180 tcaaggaaca
gcgcgacgtc gccgatcagc aggcgcgatt cgattaagtt ggtggcaacg 240
gtcagatccg agagaccttc cagcaggcac tcttcgaggg tgcgtacgct gtggcccacc
300 tccagcttga cgtcccacag cagggtgagc agttcgccga ctttttgcgc
ctggtcgtcc 360 ggcagttttt tacgactgag gatcagcaga tcgacgtctg
agagcgggtg cag 413 56 500 DNA Klebsiella sp. 56 cttaacccgc
acgctggcga aggcggccat atgggaacag aagagataga caccatcatt 60
ccggtgctgg aagagatgcg cgcaaagggg atgaacctca gcggtccgct gccggcagac
120 actctctttc agccgaaata tcttgatcat gccgatgcgg tactcgcgat
gtaccacgat 180 cagggcctgc ccgtgctaaa ataccagggc tttggccgcg
gcgtgaacat tacgctcggt 240 ttacctttta ttcgtacctc cgtcgaccac
ggcaccgcac tggaattagc gggccaggga 300 aaagcggacg tcggcagttt
tatcacggcg cttaatctcg ccatcaaaat gattgttaat 360 acccaatgaa
taatcgagtc catcagggcc atttagcccg caaacgcttc gggcagaact 420
tcctcaacga tcagtttgtg atcgacagca tcgtctcggc gattaacccg cagaaaggcc
480 aggcgatggt tgaaatcggc 500 57 473 DNA Klebsiella sp. 57
gggtctgacc ccggttctgt gcatcggtga aaccgaagcc gaaaacgaag cgggcaaaac
60 ggaagaagtt tgcgcacgtc agatcgacgc cgtgctgaaa acccagggcg
ctgccgcttt 120 cgaaggcgtg gttatcgctt acgaaccagt atgggctatc
ggtaccggca aatcagcgac 180 cccggctcag gcgcaggcgg tgcacaaatt
catccgtgac cacattgcta aagctgacgc 240 caaaatcgct gagcaagtga
tcatccagta cggcggttcc gttaacgctg gcaacgccgc 300 agagctgttc
acccagccgg acatcgacgg cgcgctggtt ggcggcgcct ccctgaaagc 360
tgacgctttc gcggtgatcg ttaaagcagc agaagcagcg aaaaaagcgt aattcgcttt
420 tcccggtggc gacacgcgac cgggttgact gacaaaacgt gggagcccgg cct 473
58 463 DNA Klebsiella sp. 58 ggtggcgcac cctggcgtcg tttgtgtaga
aattatgaat attaatacca ggaaaattcc 60 taatttttgt gtacgctctg
acgagcgcac aataaaacaa gacgaatttt tgaacaattg 120 tctttaaatt
tgttaattga attgatctgt tgttgtttaa aggtatttga atttcttttg 180
tatagatatg taaattaaca ttgaaaagcc atttcaaaaa ttaaatatat ggcgaacata
240 gctattaact tatagttaac atcttcccgg gttgcctttt gatacttcgg
gtaatatatt 300 tatttcgcac atcaaaataa ctcttttttc ttctgtttgt
tattcatggc catctattgg 360 cgaaataagg cagagtagag ggggatgtgc
ctaatatcct gcggaaggaa cgcaatgtac 420 atttacaggg aggagctgac
gagccgtttc gcgatagctt tag 463 59 526 DNA Klebsiella sp. 59
ttggtggtgt gctcgcgaag aaatttaatc tgccggtcat cgtaagtttt gttgggcttg
60 gaagagtatt ttcttctgac agcatgcctt taaaattatt gcggcagttt
actattgctg 120 catataaata tattgccagt aataagcgct gtatatttat
gtttgaacat gaccgcgaca 180 gaaaaaaact ggctaagttg gttggactcg
aagaacaaca gactattgtt attgatggtg 240 caggcattaa tccagagata
tacaaatatt ctcttgaaca ggatcacgat gtccctgttg 300 tattgtttgc
cagccgtatg ttgtggagta aaggactggg cgacttaatt gaagcgaaga 360
aaatattacg cagtaagaat attcacttta ctttgaatgt tgctggaatt ctggtcgaaa
420 atgataaaga tgcaatttcc cttcagggtc attgaaaatt ggcatcagca
aggattaatt 480 aactggttag gtcgttcgaa taatgtttgc gatcttattg agcaat
526 60 473 DNA Klebsiella sp. 60 ttacttgccc ctttttgccg aactgaaaca
aaggcccgtg ctggtgatcg gcggcggcga 60 gattgctgaa cgtaagatca
agttcctgct gcgcgcccag gcgcaggtgc aggtggtcgc 120 tgaaacgctg
tcaccggcgc tggccgatct ggctgcgcgc caggcactca gctggcgggc 180
gacggcattc agcgactcgc tggtggatga tgtctttctg gtgattgcgg ccaccgagga
240 tgaggcgctt aaccagcggg tgtttgcggc agctaacgcg cgctaccggt
tggtcaacgt 300 ggtggataac caggcgctgt gctcgtttgt tttcccttct
atcgtcgacc gttcgccgct 360 gctggtggcg atctcctcca gcggtaaagc
gccggtgttg tcgcgcattc tgcgtgaaaa 420 aatcgaagcg ctgctgccga
cgaatctcgg tcggctggcg gaatcagcaa gct 473 61 451 DNA Klebsiella sp.
61 agcagggcaa tggtggtcgg tttcataaca tttcctgatg atgaaagtca
tattaaccgg 60 cattctaaca gcagcattca gaggggcaat gattttgggc
aaccgattac gacgatcgcc 120 gcaaatgcta aaaaagggag aggggattac
cagctggcgg gcttttccgc gccgagatta 180 tccagcacgg cgcgcagcgc
caggccgtca ggaaagtgaa ggtccggggc gatctcgaac 240 agcggccaga
gcataaagcc gcggtttttc atatcgtagt gcggaacggt caggcgctcg 300
ctgttaatga cagcatcgcc aaacagcatg atatcgaggt ccagcgtgcg cggcccccag
360 cgttcggctt tgcgcactcg cccctgctgc agttcgatgc gctgagtatg
atcgagcagc 420 gtctcggggg gcagggcggt ttccagcgca a 451 62 525 DNA
Klebsiella sp. 62 ggcttaacgc cagctatgtc aacgctgcgg ttatgcggat
ttttcatgcc tctgcggcta 60 acagaaaaaa gccttatgat agctatacta
atggggcttt ttactccgtt ttgacccgat 120 tcctgaccgg cgtcagggtc
aagtcacaaa aatcatcaca attttccgtc accggcgcta 180 caatcgaccg
aagtcacaat ctcaaatcag aagagtattg ctaatgaaaa acatcaaccc 240
aacgcagacc tctgcctggc aggcattaca gaaacacttc gacgaaatga aagatgtcac
300 tatcagcgag cttttcgcca aagatagcga ccgtttttct aaattttccg
cgacgttcga 360 cgatctgatg ctggtggact tctccaaaaa ccgcatcact
gaagagacgc tggctaaact 420 gcaggatctg gcgaaagaga ctgacctggc
gggcgctatc aagtcgatgt tctcaggtga 480 gaagatcaac cgcaccgaag
accgcgcggt actgcacgtc gcgct 525 63 475 DNA Klebsiella sp. 63
tgcttcatcc gcatctcctt gaaatttatt tggtcttagg cggacggtag agcgctaata
60 gctcgtccac ctttttacgc gtaccaccgt tgctgctgat gctgcgccgc
accttcacaa 120 tatgcgtttc tgccgcgttt ttataccatt cctgcgtcag
cggcgtgcgg tggttggaaa 180 tcagcaccgg gatgcgcttt ttcatcagcg
attccgcctt ttgcgccagc agtacctgtt 240 gttccaggtt gaaactgttg
gtgtggtagg cggtaaagtt cgccgtcgcc gttagcggcg 300 catagggcgg
atcgcaatac accactgtgc ggctatccgc acgttgcatg cactcttcgt 360
aagattcgca gtaaaactcg gcgttttgcg ccttctcggc gaaatgatag agctcagctt
420 cggggaaata gggcttttta taacggccaa acggcacatt gaactcgccg cgcag
475 64 286 DNA Klebsiella sp. 64 tgtcaatgcg caatttggtt aaatatgtcg
gtattggcct gctggtgatg gggcttgccg 60 cctgcgataa cagcgattca
aaagcgccaa ccgttggcgc agcagcggag agcaatgcca 120 gcggccaggc
aatcagcctg ctggatggca agctgagctt caccctgcct gcgggcatgg 180
ccgaccagag cggcaaactg ggtacccagg cgaacaatat gcacgtctac tctgacgcta
240 ccggccagaa agcggtcatc gtcatcgtcg gcgacagcac caatga 286
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