Polynucleotides encoding polypeptides involved in the stress response to environmental changes in Methylophilus methylotrophus

Abstract

The present invention provides polypeptides and polynucleotides involved in the stress response to environmental changes in Methylophilus methylotrophus and methods of producing amino acids in microorganisms having enhanced or attenuated expression of these polypeptides and/or polynucleotides.

Description

[0001] This application is a divisional under 35 U.S.C. .sctn.120 of Ser. No. 10/375,010, filed Feb. 28, 2003, the entirety of which is incorporated by reference.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to novel polynucleotides encoding proteins involved in the stress response to environmental changes including DNA damages, phage infection and physiological stresses, derived from microorganisms belonging to methylotrophic bacteria and fragments thereof, polypeptides encoded by the polynucleotides and fragments thereof, polynucleotide arrays comprising the polynucleotides and fragments thereof.

[0004] 2. Discussion of the Background

[0005] Amino acids such as L-lysine, L-glutamic acid, L-threonine, L-leucine, L-isoleucine, L-valine and L-phenylalanine are industrially produced by fermentation by using microorganisms that belong to the genus Brevibacterium, Corynebacterium, Bacillus, Escherichia, Streptomyces, Pseudomonas, Arthrobacter, Serratia, Penicillium, Candida or the like. In order to improve the productivity of amino acids, strains of the aforementioned microorganisms that have been isolated from nature or artificial mutants thereof have been used. Various techniques have also been disclosed for enhancing activities of L-amino acid biosynthetic enzymes by using recombinant DNA techniques to increase the L-amino acid-producing ability.

[0006] L-amino acid production has been increased considerably by breeding of microorganisms such as those mentioned above and by improvements in production methods. However, in order to meet a future increase in the demand for L-amino acids, development of methods for more efficiently producing L-amino acids at lower cost are still desired.

[0007] Conventional methods for producing amino acids by fermentation using methanol, which is a raw fermentation material available in large quantities at a low cost, employ Achromobacter or Pseudomonas microorganisms (Japanese Patent Publication (Kokoku) No. 45-25273/1970), Protaminobacter microorganisms (Japanese Patent Application Laid-open (Kokai) No. 49-125590/1974), Protaminobacter or Methanomonas microorganisms (Japanese Patent Application Laid-open (Kokai) No. 50-25790/1975), Microcyclus microorganisms (Japanese Patent Application Laid-open (Kokai) No. 52-18886/1977), Methylobacillus microorganisms (Japanese Patent Application Laid-open (Kokai) No. 4-91793/1992), Bacillus microorganisms (Japanese Patent Application Laid-open (Kokai) No. 3-505284/1991) and others.

[0008] However, only a few methods have been described for producing L-amino acids using Methylophilus bacteria in conjunction with recombinant DNA technology. Although methods described in EP 0 035 831 A, EP 0 037 273 A and EP 0 066 994 A have been described as methods for transforming Methylophilus bacteria using recombinant DNA, applying recombinant DNA techniques to improvement of amino acid productivity of Methylophilus bacteria has not been described. Only WO-00/61723 and WO-02/38777 disclose the improved production of lysine and phenylalanine, respectively, using genes involved in biosynthesis of each respective amino acid. In particular, WO-00/61723 discloses the ask gene, the dapA gene, the dapB gene, and the lysA gene, which encode aspartkinase, dihydrodipicolinate synthase, dihydrodipicolinate reductase, and diaminopimelinate decarboxylase, respectively. WO-02/38777 discloses the aroG gene and the pheA gene, which encode 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase and bi-functional enzyme of chorismate mutase and prephenate dehydratase, respectively.

[0009] Therefore, prior to the present invention genes isolated from Methylophilus bacteria that are involved in the stress response to environmental changes and which can be used to improve the yield of amino acids in cultured microorganisms remain elusive and undisclosed.

SUMMARY OF THE INVENTION

[0010] An object of the present invention is to provide novel measures for the improved production of amino acids or an amino acid, where these amino acids include asparagine, threonine, serine, glutamate, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, tryptophan, arginine and the salts thereof. In a preferred embodiment the amino acids are L-amino acids.

[0011] Such a process includes bacteria, which express a protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34.

[0012] In one embodiment the polypeptides are encoded by a polynucleotide selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33. In another embodiment the polypeptides are encoded by other polynucleotides which have substantial identity to the herein described polynucleotides or those which hybridize under stringent conditions.

[0013] Another object of the invention is to provide polynucleotide sequences selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:1, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33; as well as those polynucleotides that have substantial identity to these nucleotide sequences, preferably at least 95% identity.

[0014] Another object of the invention is to provide isolated polypeptides having a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34; as well as those polypeptides that have substantial identity to these amino acid sequences, preferably at least 95% identity.

[0015] A further object of the invention is a method for producing a protein or proteins by culturing host cells containing the herein described polynucleotides under conditions and for a time suitable for expression of the protein and collecting the protein produced thereby.

[0016] Another object is the use of host cells having the polynucleotides described herein to produce amino acids, as well as the use of such isolated polypeptides in the production of amino acids.

[0017] Other objects of the invention include methods of detecting nucleic acid sequences homologous to at least one of: SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, particularly nucleic acid sequences encoding polypeptides that herein described proteins or polypeptides and methods of making nucleic acids encoding such polypeptides.

[0018] The above objects highlight certain aspects of the invention. Additional objects, aspects and embodiments of the invention are found in the following detailed description of the invention.

DETAILED DESCRIPTION OF THE INVENTION

[0019] 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 of molecular biology. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described herein. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0020] Reference is made to standard textbooks of molecular biology that contain definitions and methods and means for carrying out basic techniques, encompassed by the present invention. See, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Third Edition, Cold Spring Harbor Laboratory Press, New York (2001), Current Protocols in Molecular Biology, Ausebel et al (eds.), John Wiley & Sons, New York (2001) and the various references cited therein.

[0021] Methylophilus methylotrophus (M. methylotrophus) is a gram negative ribulose monophosphate cycle methanol-utilizer, which can be used for the large-scale production of a variety of fine chemicals including amino acids, nucleic acids, vitamins, saccharides, and so on. The polynucleotides of this invention, therefore, can be used to identify microorganisms, which can be used to produce fine chemicals, for example, by fermentative processes. Modulation of the expression of the polynucleotides encoding proteins in the stress response to environmental changes of the present invention, can be used to modulate the production of one or more fine chemicals from a microorganism (e.g., to improve the yield of production of one or more fine chemicals from Methylophilus or Methylbacillus species).

[0022] The proteins encoded by the polynucleotides of the present invention are capable of, for example, performing a function involved in the stress response to environmental changes in M. methylotrophus, such as, DNA damages, phage infection and several physiological stresses.

[0023] Given the availability of cloning vectors used in M. methylotrophus, such as those disclosed in Methane and Methanol Utilizers, Plenum Press, New York (1992) edited by J. Colin Murrell and Howard Dalton, the nucleic acid molecules of the present invention may be used in the genetic engineering of this organism to make it better or more efficient producer of one or more fine chemicals.

[0024] There are a number of mechanisms by which the alteration of a protein of the present invention may affect the yield, production, and/or efficiency of production of a fine chemical from M. methylotrophus bacteria, which have the altered protein incorporated.

[0025] Improving the ability of the cell to produce integration host factors (e.g., by manipulating the genes expression or the efficiency of translation of the mRNAs), one could efficiently transfer transposon, such as Mu-phage, in the host cell in order to amplify the desired gene-cassette on the chromosome using this phage-function. For example, one can place the important genes for amino acid synthesis onto the gene-cassette, and the efficiency of the transposition or amplification of the gene-cassette derived from transposon, such as Mu-phage, could be increased by the function of integration host factors. In addition, several recombinant proteins, RecR, RecN, and RecO, could facilitate the reactions. The rapid DNA manipulation may save the cost associated with breeding of the microorganism during the production of the desired fine-chemicals. On the other hand, the inactivations of these genes might give the host cells the tolerance to some bacterial phages. The property may be advantageous for large-scale fermentation of the microorganism during the production of the desired fine-chemicals.

[0026] Improving the ability of the cell to tolerate some stresses could increase the yield or productivity of desired fine chemicals, since the disturbed physiological conditions for the over-production may induce mutations in the producer cells resulting in low-productivity. Therefore, the improvement of stress tolerance may be favorable to the microorganism for the over-production of the desired fine-chemicals, and may allow the microorganism to sustain high-level productivity. Those candidate genes are RNA polymerase sigma factor D (rpoD), RNA polymerase sigma factor E (rpoE), RNA polymerase sigma factor H (rpoH), RNA polymerase sigma factor N (rpoN), Chaperone protein DnaK (dnaK), Chaperone protein DnaJ (dnaJ), Chaperone protein HscA (hscA), Chaperone protein HscB (hscB), Chaperone protein GroEL1 (groEL1), Chaperone protein GroES1 (groES1), Chaperone protein GroEL2 (groEL2), and Chaperone protein GroES2 (groES2).

[0027] "L-amino acids" or "amino acids" as used herein means one or more amino acids, including their salts, preferably chosen from the following: L-asparagine, L-threonine. L-serine, L-glutamate, L-glycine, L-alanine, L-cysteine, L-valine, L-methionine, L-isoleucine, L-leucine, L-tyrosine, L-phenylalanine, L-histidine, L-lysine, L-tryptophan and L-arginine.

[0028] "Isolated" as used herein means separated out of its natural environment.

[0029] "Substantial identity" as used herein refers to polynucleotides and polypeptides which are at least 70%, preferably at least 80% and more preferably at least 90% to 95% identical to the polynucleotides and polypeptides, respectively, according to the present invention.

[0030] "Polynucleotide" as used herein relates to polyribonucleotides and polydeoxyribonucleotides, it being possible for these to be non-modified RNA or DNA or modified RNA or DNA.

[0031] "Polypeptides" as used herein are understood to mean peptides or proteins which comprise two or more amino acids bonded via peptide bonds. In particular, the term refers to polypeptides which are at least 70%, preferably at least 80% and more preferably at least 90% to 95% identical to the polypeptides according to the present invention. Included within the scope of the present invention are polypeptide fragments of the polypeptides having a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34 or those which are identical to those described herein.

[0032] "Polynucleotides which encode the polypeptide" of the invention as used herein is understood to mean the sequences exemplified in this application as well as those sequences which have substantial identity to the nucleic acid sequences at least one of: SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33 and which encode a molecule having one or more of the bioactivities of the associated gene products. Preferably, such polynucleotides are those which are at least 70%, preferably at least 80% and more preferably at least 90% to 95% identical to the nucleic acid sequences at least one of: SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33.

[0033] Polynucleotides according to the invention may be employed as probes to isolate and/or identify RNA, cDNA and DNA molecules, e.g., full-length genes or polynucleotides which code for the polypeptides described herein. Likewise, the probes can be employed to isolate nucleic acids, polynucleotides or genes which have a high sequence similarity or identity with the polynucleotides of the invention.

[0034] Polynucleotides of the invention may also be used to design primers useful for the polymerase chain reaction to amplify, identify and/or isolate full-length DNA, RNA or other polynucleotides with high sequence homology or identity to the polynucleotides of the invention, as well as, polynucleotides that encode the polypeptides of the invention. Preferably, probes or primers are at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleotides in length. Oligonucleotides with a length of at least 35, 40, 45, 50, 100, 150, 200, 250 or 300 nucleotides may also be used.

[0035] Methods of DNA sequencing are described inter alia by Sanger et al. (Proceedings of the National Academy of Sciences of the United States of America USA, 74:5463-5467, (1977)).

[0036] A person skilled in the art will find instructions for amplification of DNA sequences with the aid of the polymerase chain reaction (PCR) inter alia in the handbook by Gait: Oligonucleotide Synthesis: A Practical Approach (IRL Press, Oxford, UK, 1984) and in Newton and Graham: PCR 2.sup.nd Edition (Springer Verlag, New York, 1997).

[0037] Additionally, methods employing DNA chips, microarrays or similar recombinant DNA technology that enables high throughput screening of DNA and polynucleotides that encode the herein described proteins or polynucleotides with high sequence homology or identity to the polynucleotides described herein. Such methods are known in the art and are described, for example, in Current Protocols in Molecular Biology, Ausebel et al (eds), John Wiley and Sons, Inc. New York (2000).

[0038] The polynucleotides and polypeptides of the present invention are involved in the stress response to environmental changes in M. methylotrophus. By way of example, the present inventors provide the following cited references (each of which are incorporated herein by reference) demonstrating that assays to assess the enzymatic activity of the polypeptides of the present invention are known and, as such, determination of whether a sequence falls within the scope of the present claims may be readily ascertained. These polynucleotides and polypeptides include: [0039] 1. Recombinant protein RecR comprises the amino acid sequence of SEQ ID NO:2 and is encoded by the recR gene which comprises the polynucleotide SEQ ID NO:1 (According to Pelaez, A. I. et. al. Mol. Gen. Genet. (2001) 265:663-672, the recR deficient mutant is more sensitive to DNA damage caused by UV irradiation or meythylmethane sulfonate than the wild type); [0040] 2. Recombinant protein RecN comprises the amino acid sequence of SEQ ID NO:4 and is encoded by the recN gene which comprises the polynucleotide SEQ ID NO:3 (The recN disruption mutant shows sensitivity to mitomycin C and ionizing radiation (Picksley, S. M. et. al. Mol. Gen. Genet. (1984) 195:267-274)); [0041] 3. Recombinant protein RecO comprises the amino acid sequence of SEQ ID NO:6 and is encoded by the recO gene which comprises the polynucleotide SEQ ID NO:5 (The RecO activity can be measured as described by Luisi-DeLuca, C. and Kolodner, R (J. Mol. Biol. (1994) 236:124-138)); [0042] 4. Integration host factor alpha-subunit enzyme comprises the amino acid sequence of SEQ ID NO:8 and is encoded by the infA gene which comprises the polynucleotide SEQ ID NO:7 (The activity of integration host factor can be measured through complementation of E. coli ifhA and ihfB mutant (Club, R. et. al., J. Bacteriol. (1996) 178:6319-6326)); [0043] 5. Integration host factor beta-subunit enzyme comprises the amino acid sequence of SEQ ID NO:10 and is encoded by a infB gene which comprises the polynucleotide SEQ ID NO:9 (The activity of integration host factor can be measured through complementation of E. coli ifhA and ihfB mutant (Club, R. et. al., J. Bacteriol. (1996) 178:6319-6326)); [0044] 6. RNA polymerase sigma factor D comprises the amino acid sequence of SEQ ID NO:12 and is encoded by a rpoD gene which comprises the polynucleotide SEQ ID NO:11 (The activity of RpoD can be assayed in combination with DNA dependent RNA polymerase core enzyme (Gribskov, M. and Burgess, R. R. Gene (1983) 26:109-118)); [0045] 7. RNA polymerase sigma factor E comprises the amino acid sequence of SEQ ID NO:14 and is encoded by a rpoE gene which comprises the polynucleotide SEQ ID NO:13 (Rouviere, P. E. et. al. EMBO J. (1995) 14:1032-1042 and Raina, S. et. al. EMBO J. (1995) 14:1043-1055); [0046] 8. RNA polymerase sigma factor H comprises the amino acid sequence of SEQ ID NO:16 and is encoded by a rpoH gene comprising SEQ ID NO:15 (Grossman, A. D. et. al. Cell (1984) 38:383-390); [0047] 9. RNA polymerase sigma factor N comprises the amino acid sequence of SEQ ID NO:18 and is encoded by a rpoN gene comprising SEQ ID NO:17 (Cannon, W. et. al. Mol. Microbiol. (1996) 21:233-245); [0048] 10. Chaperone protein DnaK comprises the amino acid sequence of SEQ ID NO:20 and is encoded by a dnaK gene comprising SEQ ID NO:19 (Zylicz, M. and Georgopoulos, C. J. Biol. Chem. (1984) 259:8820-8825); [0049] 11. Chaperone protein DnaJ comprises the amino acid sequence of SEQ ID NO:22 and is encoded by a dnaJ gene comprising SEQ ID NO:21 (Zylicz, M. et. al. J. Biol. Chem. (1985) 260:7591-7598); [0050] 12. Chaperone protein HscA comprises the amino acid sequence of SEQ ID NO:24 and is encoded by a hscA gene comprising SEQ ID NO:23 (The activity of chaperone protein HscA and HscB can be assayed as ATP hydrolysis activity (Vickery, L. E., Protein Sci. (1997) 6:1047-1056) and as inhibitory activity to protein denaturation (Silberg, J. J., J. Bacteriol. (1998) 180:6617-6624)); [0051] 13. Chaperone protein HscB comprises the amino acid sequence of SEQ ID NO:26 and is encoded by a hscB gene comprising SEQ ID NO:25 (The activity of chaperone protein HscA and HscB can be assayed as ATP hydrolysis activity (Vickery, L. E., Protein Sci. (1997) 6:1047-1056) and as inhibitory activity to protein denaturation (Silberg, J. J., J. Bacteriol. (1998) 180:6617-6624)); [0052] 14. Chaperone protein GroEL1 comprises the amino acid sequence of SEQ ID NO:28 and is encoded by a groEL1 gene comprising SEQ ID NO:27 (The activity of GroEL can be detected as ATP hydrolysis activity (Terlesky, K. C. and Tabita, F. R., Biochemistry (1991) 30:8181-8186)); [0053] 15. Chaperone protein GroES1 comprises the amino acid sequence of SEQ ID NO:30 and is encoded by a groES1 gene comprising SEQ ID NO:29 (The activity of GroES can be detected as inhibitory activity to the GroEL ATP hydrolysis activity (Terlesky, K. C. and Tabita, F. R., Biochemistry (1991) 30:8181-8186)); [0054] 16. Chaperone protein GroEL2 comprises the amino acid sequence of SEQ ID NO:32 and is encoded by a groEL2 gene comprising SEQ ID NO:31 (The activity of GroEL can be detected as ATP hydrolysis activity (Terlesky, K. C. and Tabita, F. R., Biochemistry (1991) 30:8181-8186)); [0055] 17. Chaperone protein GroES2 comprises the amino acid sequence of SEQ ID NO:34 and is encoded by a groES2 gene comprising SEQ ID NO:33 (The activity of GroES can be detected as inhibitory activity to the GroEL ATP hydrolysis activity (Terlesky, K. C. and Tabita, F. R., Biochemistry (1991) 30:8181-8186)).

[0056] The terms "stringent conditions" or "stringent hybridization conditions" includes reference to conditions under which a polynucleotide will hybridize to its target sequence, to a detectably greater degree than other sequences (e.g., at least 2-fold over background). Stringent conditions are sequence-dependent and will be different in different circumstances. By controlling the stringency of the hybridization and/or washing conditions, target sequences can be identified which are 100% complementary to the probe (homologous probing). Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of similarity are detected (heterologous probing).

[0057] Typically, stringent conditions will be those in which the salt concentration is less than approximately 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30.degree. C. for short probes (e.g., 10 to 50 nucleotides) and at least about 60.degree. C. for long probes (e.g., greater than 50 nucleotides). Stringent conditions also may be achieved with the addition of destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization with a buffer solution of 30 to 35% formamide, 1 M NaCl, 1% SDS (w/v; sodium dodecyl sulphate) at 37.degree. C., and a wash in 1.times. to 2.times.SSC (20.times.SSC=3.0 M NaCl/0.3 M trisodium citrate) at 50 to 55.degree. Exemplary moderate stringency conditions include hybridization in 40 to 45% formamide, 1 M NaCl, 1% SDS at 37.degree. C., and a wash in 0.5.times. to 1.times.SSC at 55 to 60.degree. C. Exemplary high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37.degree. C., and a wash in 0.1.times.SSC at 60 to 65.degree. C.

[0058] Specificity is typically the function of post-hybridization washes, the critical factors being the ionic strength and temperature of the final wash solution. For DNA--DNA hybrids, the Tm can be approximated from the equation of Meinkoth and Wahl (Anal. Biochem., 138:267-284, 1984): Tm=81.5.degree. C.+16.6 (log M)+0.41 (% GC)-0.61 (% form)-500/L; where M is the molarity of monovalent cations, % GC is the percentage of guanosine and cytosine nucleotides in the DNA, % form is the percentage of formamide in the hybridization solution, and L is the length of the hybrid in base pairs. The Tm is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. Tm is reduced by about 1.degree. C. for each 1% of mismatching; thus, Tm, hybridization and/or wash conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences with approximately 90% identity are sought, the Tm can be decreased 10.degree. C.

[0059] Generally, stringent conditions are selected to be about 5.degree. C. lower than the thermal melting point (Tm) for the specific sequence and its complement at a defined ionic strength and pH. However, severely stringent conditions can utilize hybridization and/or wash at 1, 2, 3, or 4.degree. C. lower than the thermal melting point (Tm); moderately stringent conditions can utilize a hybridization and/or wash at 6, 7, 8, 9, or 10.degree. C. lower than the thermal melting point (Tm); low stringency conditions can utilize a hybridization and/or wash at 11, 12, 13, 14, 15, or 20.degree. C. lower than the thermal melting point (Tm). Using the equation, hybridization and wash compositions, and desired Tm, those of ordinary skill will understand that variations in the stringency of hybridization and/or wash solutions are inherently described. If the desired degree of mismatching results in a Tm of less than 45.degree. C. (aqueous solution) or 32.degree. C. (formamide solution), it is preferred to increase the SSC concentration so that a higher temperature can be used. An extensive guide to the hybridization of nucleic acids is found in Tijssen, Laboratory Techniques in Biochemistry and Molecular Biology--Hybridization with Nucleic Acid Probes, Part 1, Chapter 2 "Overview of principles of hybridization and the strategy of nucleic acid probe assays", Elsevier, New York (1993); and Current Protocols in Molecular Biology, Chapter 2, Ausubel, et al., Eds., Greene Publishing and Wiley-Interscience, New York (1995).

[0060] Stringent hybridization conditions are understood to mean those conditions where hybridization, either in solution or on a solid support, occur between two polynucleotide molecules which are 70% to 100% homologous in nucleotide sequence which include 75%, 80%, 85%, 90%, 95%, 98% and all values and subranges therebetween.

[0061] Homology, sequence similarity or sequence identity of nucleotide or amino acid sequences may be determined conventionally by using known software or computer programs. To find the best segment of identity or similarity of sequences, BLAST (Altschul et al (1990) J. Mol. Biol. 215:403-410 and Lipman et al (1990) J. Mol. Biol. 215:403-410), FASTA (Lipman et al (1985) Science 227:1435-1441), or Smith and Waterman (Smith and Waterman (1981) J. Mol. Biol. 147:195-197) homology search programs can be used. To perform global alignments, sequence alignment programs such as the CLUSTAL W (Thompson et al (1994) Nucleic Acids Research 22:4673-4680) can be used.

[0062] The present invention also provides processes for preparing amino acids using bacteria that comprise at least one polynucleotide whose expression is enhanced or attenuated. Likewise, the invention also provides processes for preparing amino acids using bacteria that comprise at least on polypeptide whose activity is enhanced or attenuated. Preferably, a bacterial cell with enhanced or attenuated expression of one or more of the polypeptides and/or polynucleotides described herein will improve amino acid yield at least 1% compared to a bacterial strain not having the enhanced or attenuated expression. For the production of amino acids the M. methylotrophus polynucleotides described herein may be used to target expression, either by disruption to turn off or increase or enhance the expression or relative activity of the polypeptide enzymes encoded therein.

[0063] The term "enhancement" as used herein means increasing intracellular activity of one or more polypeptides in the bacterial cell, which in turn are encoded by the corresponding polynucleotides described herein. To facilitate such an increase, the copy number of the genes corresponding to the polynucleotides described herein may be increased. Alternatively, a strong and/or inducible promoter may be used to direct the expression of the polynucleotide, the polynucleotide being expressed either as a transient expression vehicle or homologously or heterologously incorporated into the bacterial genome. In another embodiment, the promoter, regulatory region and/or the ribosome binding site upstream of the gene can be altered to achieve the over-expression. The expression may also be enhanced by increasing the relative half-life of the messenger RNA.

[0064] In another embodiment, the enzymatic activity of the polypeptide itself may be increased by employing one or more mutations in the polypeptide amino acid sequence, which increases the activity. For example, altering the relative Km of the polypeptide with its corresponding substrate will result in enhanced activity. Likewise, the relative half-life of the polypeptide may be increased.

[0065] In either scenario, that being enhanced gene expression or enhanced enzymatic activity, the enhancement may be achieved by altering the composition of the cell culture media and/or methods used for culturing.

[0066] "Enhanced expression" or "enhanced activity" as used herein means an increase of at least 10%, 25%, 50%, 75%, 100%, 150%, 200%, 300%, 400% or 500% compared to a wild-type protein, polynucleotide, gene; or the activity and/or the concentration of the protein present before the polynucleotides or polypeptides are enhanced.

[0067] The term "attenuation" as used herein means a reduction or elimination of the intracellular activity of the polypeptides in a bacterial cell that are encoded by the corresponding polynucleotide. To facilitate such a reduction or elimination, the copy number of the genes corresponding to the polynucleotides described herein may be decreased or removed. Alternatively, a weak and/or inducible promoter may used to direct the expression of the polynucleotide, the polynucleotide being expressed either as a transient expression vehicle or homologously or heterologously incorporated into the bacterial genome. For example, the endogenous promoter or regulatory region of the gene corresponding to the isolated polynucleotides described herein may be replaced with the aforementioned weak and/or inducible promoter. Alternatively, the promoter or regulatory region may be removed. The expression may also be attenuated by decreasing the relative half-life of the messenger RNA.

[0068] In another embodiment, the enzymatic activity of the polypeptide itself may be decreased or deleted by employing one or more mutations in the polypeptide amino acid sequence, which decreases the activity or removes any detectable activity. For example, altering the relative Kd of the polypeptide with its corresponding substrate will result in attenuated activity. Likewise, a decrease in the relative half-life of the polypeptide will result in attenuated activity.

[0069] By attenuation measures, the activity or concentration of the corresponding protein is in general reduced to 0 to 75%, 0 to 50%, 0 to 25%, 0 to 10% or 0 to 5% of the activity or concentration of the wild-type protein or of the activity or concentration of the protein in the starting microorganism.

[0070] Suitable vectors for carrying M. methylotrophus polynucleotides include those vectors which can direct expression of the gene in bacterial cells as known in the art. One embodiment of the present invention is whereby the vectors contain an inducible or otherwise regulated expression system whereby the M. methylotrophus polynucleotides may be expressed under certain conditions and not expressed under other conditions. Furthermore, in another embodiment of the invention, the M. methylotrophus polynucleotides can be constitutively expressed. Examples of such vectors and suitable cells in which they can be introduced are described in Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd Ed., Cold Spring Harbor Laboratory Press,, Cold Spring Harbor, N.Y. (2001) and Current Protocols in Molecular Biology, Ausebel et al, (Eds.), John Wiley and Sons, Inc., New York, 2000.

[0071] Methods of introducing M. methylotrophus polynucleotides or vectors containing the M. methylotrophus polynucleotides include electroporation, conjugation, calcium-mediated transfection, infection with bacteriophage and other methods known in the art. These and other methods are described in Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2001) and Current Protocols in Molecular Biology, Ausebel et al, (Eds.), John Wiley and Sons, Inc., New York (2000).

[0072] The microorganisms that can be used in the present invention should have the ability to produce amino acids, preferably L-amino acids, from a suitable carbon source, preferably carbon sources such as methanol, glucose, sucrose, lactose, fructose, maltose, molasses, starch, cellulose glycerol or ethanol. The microorganisms can be Methylophilus bacteria, preferably Methylophilus methylotrophus.

[0073] Suitable culture conditions for the growth and/or production of M. methylotrophus polynucleotides are dependent on the cell type used. Likewise, culturing cells that contain attenuated or enhanced expression of the M. methylotrophus polynucleotides or polypeptides, as described herein, may be cultured in accordance with methods known in the art. Examples of culture conditions for various cells is described in Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2001); Current Protocols in Molecular Biology, Ausebel et al, (Eds.), John Wiley and Sons, Inc., 2000; and Cells: A Laboratory Manual (Vols. 1-3), Spector et al, (Eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1988).

[0074] Following culturing the polypeptide or protein products, which are encoded by the M. methylotrophus polynucleotides, may be purified using known methods of protein purification. These methods include high performance liquid chromatography (HPLC), ion-exchange chromatography, size exclusion chromatography; affinity separations using materials such as beads with exposed heparin, metals, or lipids; or other approaches known to those skilled in the art. These and other methods of protein purification are disclosed in Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2001); Current Protocols in Molecular Biology, Ausebel et al, eds., John Wiley and Sons, Inc., 2000 and Protein Purification, Scopes and Cantor, (Eds.), Springer-Verlag, (1994). Likewise, the amino acids produced may be purified by methods known in the art using similar chromatography devices.

[0075] The invention also provides antibodies that bind to the polypeptides of the present invention. Antibodies binding to the polypeptides can be either monoclonal or polyclonal, preferably the antibodies are monoclonal. Methods for obtaining antibodies that bind to the polypeptides are known in the art and are described, for example, in Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1988).

[0076] Having generally described this invention, a further understanding can be obtained by reference to certain specific examples which are provided herein for purposes of illustration only, and are not intended to be limiting unless otherwise specified.

EXAMPLES

[0077] Whole genome sequencing using random shotgun method is described in Fleischman R. D. et. al. (1995) Science, 269: 496-512.

Example 1

Construction of Genomic Libraries of Methylophilus methylotrophus

[0078] M. methylotrophus AS1 was cultured at 30.degree. C. in the 121 medium described in the Catalogue of Strains (The National Collections of Industrial and Marine Bacteria Ltd., 1994).

[0079] Cells were collected by centrifugation. Genomic DNA was isolated using the Genome-tip system (Qiagen K. K., Tokyo, Japan). The genomic DNA was sheared and fragmentized by sonication. The resultant fragments in the 1- to 2-kb size range were purified by gel electrophoresis through 1% low-melting agarose gel, followed by recovery using the Wizard DNA purification kit (Promega KK, Tokyo, Japan). The recovered fragments were ligated to the high-copy number vector pUC118 treated by HincII and bacterial alkaline phosphatase (Takara Shuzo, Kyoto, Japan), and this was designated pUC 118 library.

[0080] For larger fragments (9- to 11-kb in size), the genomic DNA was partially digested by restriction endonuclease Sau3AI, followed by 0.6% agarose gel electrophoresis. The DNA fragments corresponding 9-kb to 11-kb in size were excised from gel and were recovered using the DNACELL (Daiichi Pure Chemicals, Tokyo, Japan). The recovered fragments were ligated into the low-coy number vector pMW118 (Nippon Gene, Toyama, Japan), which is a derivative of the pSC101 (Bernaidi, A. and Bernardi, F. (1984) Nucleic Acids Res. 12, 9415-9426). This library composed of large DNA fragments was designated pMW118 library.

[0081] General DNA manipulation was performed according to previously described methods (Sambrook et. al. (1989) "Molecular Cloning: A Laboratory Manual/Second Edition", Cold Spring Harbor Laboratory Press).

Example 2

DNA Sequencing and Sequence Assembly

[0082] The pUC118 library were transformed into Escherichia coli DH5.alpha. and plated on Luria-Bertani medium containing 100 .mu.g/ml ampicillin and 40 .mu.g/ml 5-bromo-4-chloro-3-indolyl-.alpha.-D-galactoside (X-Gal). The white colonies were picked up and cultured in Luria-Bertani medium containing 100 .mu.g/ml ampicillin. The individual colony was cultured in the well of the 96 deep-well plates, and the plasmids were isolated using QIAprep Turbo Kit (Qiagen). The DNA fragments inserted into pUC118 were sequenced using a M13 reverse primer. The shotgun sequencing was performed with the BigDye terminators and 3700 DNA analyzer (Applied Biosystems Japan, Tokyo, Japan). Approximately 50,000 samples from pUC118 library corresponding to coverage of approximately 8-fold to the genome size were analyzed and the sequences were assembled by Phred/Phrap software (CodonCode, MA, USA). This assembly treatment yielded 60 contigs with more than 5 kb in size.

[0083] As for pMW118 library, 2,000 clones corresponding to coverage of approximately 5-fold were sequenced using both M13 forward and reverse primers. The end-sequence data were analyzed and the linking clones between contigs were selected from pMW118 library. The inserted fragments of selected clones were amplified by the polymerase chain reaction (PCR) using LA Taq polymerase (Takara Shuzo) and M. methylotrophus genomic DNA as a template. These products of PCR were entirely sequenced as described in Example 1, and the gap DNA sequences between contigs were determined. By the additional sequence information, the Phrap assembly software reduced the number of contigs with more than 5 kb in size to 24. Then the 48 DNA primers with sequences complementary to the end-sequences of the 24 contigs were prepared. All possible pairwise combination of the primers were tested by PCR to amplify the DNA fragments of M. methylotrophus genomic DNA. The amplified products were sequenced directly. In several cases, the additional primers complementary to different sequences at the end of the contig were used. This strategy could close all of the remaining physical gaps and resulted in a single circular contig. Several regions that had been sequenced in only one direction and had postulated secondary structure were confirmed. By this research, the genome of M. methylotrophus was found to be a single circular with the size of 2,869,603 bases and GC content of 49.6%.

Example 3

Sequence Analysis and Annotation

[0084] Sequence analysis and annotation was managed using the Genome Gambler software (Sakiyama, T. et. al. (2000) Biosci. Biotechnol. Biochem. 64: 670-673). All open reading frames of more than 150 bp in length were extracted and the translated amino acid sequences were searched against non-redundant protein sequences in GenBank using the BLAST program (Altschul, S. F. et. al. (1990) J. Mol. Biol. 215, 403-410). Of putative polynucleotide encoding sequences with significant similarities to the sequences in public databases (BLASTP scores of more than 100), the genes involved in biosynthesis of amino acids were selected. Start codons (AUG or GUG) were putatively identified by similarity of the genes and their proximity to the ribosome binding sequences (Shine, J. and Dalgarno, L. (1975) Eur. J. Biochem. 57: 221-230). Careful assignment of gene function resulted in the identification of the recombination protein genes (recR, recN, and recO), integration host factor genes (infA, and infB), and RNA polymerase sigma factor genes (rpoD, rpoE, rpoH, and rpoN). Also, chaperone proteins genes (dank, dnaJ, hscA, hscB, groEL, and groES) were identified. Two pairs of chaperone protein genes, dnaK-dnaJ and hscA-hscB were found in a putative operon, respectively. On the other hand, 2 copies chaperone protein genes groEL-groES were found in the genome. Therefore we designated them as groEL1-groES1 and groEL2-groES2.

[0085] Obviously, numerous modifications and variations on the present invention are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described herein.

[0086] While the invention has been described in detail with reference to preferred embodiments thereof, it will be apparent to one skilled in the art that various changes can be made, and equivalents employed, without departing from the scope of the invention. Each of the aforementioned documents is incorporated by reference herein in its entirety.

Sequence CWU 1

1

34 1 543 DNA Methylophilus methylotrophus CDS (1)..(543) 1 gtg ggt ccc aag tcg tcc aca cgc atg gct tat cac ctc ctg cag cgg 48 Val Gly Pro Lys Ser Ser Thr Arg Met Ala Tyr His Leu Leu Gln Arg 1 5 10 15 gac aga gaa ggt gcg caa aaa ctg gct gcg agt ttg cag cag gcg ctg 96 Asp Arg Glu Gly Ala Gln Lys Leu Ala Ala Ser Leu Gln Gln Ala Leu 20 25 30 gac aag ctc gct cat tgt gat tat tgc aat aac ttc agc gaa cac cag 144 Asp Lys Leu Ala His Cys Asp Tyr Cys Asn Asn Phe Ser Glu His Gln 35 40 45 gta tgt agt gtg tgc ctg gaa acc agc cgt gac aaa tca acg cta tgc 192 Val Cys Ser Val Cys Leu Glu Thr Ser Arg Asp Lys Ser Thr Leu Cys 50 55 60 gtg gtc gaa atg cct acc gac ctc atg atg ctg gaa aac acc cgc agt 240 Val Val Glu Met Pro Thr Asp Leu Met Met Leu Glu Asn Thr Arg Ser 65 70 75 80 tac caa ggt caa tat ttt gta ctg atg ggc aaa cta tcg ccc atg gat 288 Tyr Gln Gly Gln Tyr Phe Val Leu Met Gly Lys Leu Ser Pro Met Asp 85 90 95 ggt ata gga ccc aaa gat att cat ctc gat aaa ctg ctc aag cgc gcg 336 Gly Ile Gly Pro Lys Asp Ile His Leu Asp Lys Leu Leu Lys Arg Ala 100 105 110 caa gat acc gcc att cat gaa gtc atc ctg gcc acc aat ttc acc tcc 384 Gln Asp Thr Ala Ile His Glu Val Ile Leu Ala Thr Asn Phe Thr Ser 115 120 125 gag ggc gaa gcc aca gcc cac tat ata gga cag atg ctc aaa tca cgt 432 Glu Gly Glu Ala Thr Ala His Tyr Ile Gly Gln Met Leu Lys Ser Arg 130 135 140 ggc atc cgt gtg agc cgg att gcc cgc ggc tta cct atg ggt gga gaa 480 Gly Ile Arg Val Ser Arg Ile Ala Arg Gly Leu Pro Met Gly Gly Glu 145 150 155 160 att gag tac gtc gac agc ggc acc ctc tcg aca gcc ctg ctg gaa cgc 528 Ile Glu Tyr Val Asp Ser Gly Thr Leu Ser Thr Ala Leu Leu Glu Arg 165 170 175 aag gtt tta act tag 543 Lys Val Leu Thr 180 2 180 PRT Methylophilus methylotrophus 2 Val Gly Pro Lys Ser Ser Thr Arg Met Ala Tyr His Leu Leu Gln Arg 1 5 10 15 Asp Arg Glu Gly Ala Gln Lys Leu Ala Ala Ser Leu Gln Gln Ala Leu 20 25 30 Asp Lys Leu Ala His Cys Asp Tyr Cys Asn Asn Phe Ser Glu His Gln 35 40 45 Val Cys Ser Val Cys Leu Glu Thr Ser Arg Asp Lys Ser Thr Leu Cys 50 55 60 Val Val Glu Met Pro Thr Asp Leu Met Met Leu Glu Asn Thr Arg Ser 65 70 75 80 Tyr Gln Gly Gln Tyr Phe Val Leu Met Gly Lys Leu Ser Pro Met Asp 85 90 95 Gly Ile Gly Pro Lys Asp Ile His Leu Asp Lys Leu Leu Lys Arg Ala 100 105 110 Gln Asp Thr Ala Ile His Glu Val Ile Leu Ala Thr Asn Phe Thr Ser 115 120 125 Glu Gly Glu Ala Thr Ala His Tyr Ile Gly Gln Met Leu Lys Ser Arg 130 135 140 Gly Ile Arg Val Ser Arg Ile Ala Arg Gly Leu Pro Met Gly Gly Glu 145 150 155 160 Ile Glu Tyr Val Asp Ser Gly Thr Leu Ser Thr Ala Leu Leu Glu Arg 165 170 175 Lys Val Leu Thr 180 3 1659 DNA Methylophilus methylotrophus CDS (1)..(1659) 3 atg cta caa gcc ctt tct atc cga gac ttt gtc att gtt gat acg ctg 48 Met Leu Gln Ala Leu Ser Ile Arg Asp Phe Val Ile Val Asp Thr Leu 1 5 10 15 gaa ctt gag ttc tca gca gga tac acc gca ctc act ggt gag acg ggt 96 Glu Leu Glu Phe Ser Ala Gly Tyr Thr Ala Leu Thr Gly Glu Thr Gly 20 25 30 gcg ggt aaa tcc atc ctc att gat gcc ctg tct tta agc ctg ggc gca 144 Ala Gly Lys Ser Ile Leu Ile Asp Ala Leu Ser Leu Ser Leu Gly Ala 35 40 45 cgt aac gag ggg gat gtg acg cgc aag ggc tgt gaa aag gcg gaa atc 192 Arg Asn Glu Gly Asp Val Thr Arg Lys Gly Cys Glu Lys Ala Glu Ile 50 55 60 tca acg act ttt gat att gcg gat aat ctg gca gcc cgg gat tgg cta 240 Ser Thr Thr Phe Asp Ile Ala Asp Asn Leu Ala Ala Arg Asp Trp Leu 65 70 75 80 cag gca caa gaa atc gat att gag gat gcc ctg gta ttg cgc cgt gtg 288 Gln Ala Gln Glu Ile Asp Ile Glu Asp Ala Leu Val Leu Arg Arg Val 85 90 95 att tat gcc gat ggc cgt agc cgc gcc ttt att aat ggt gcc tcg gca 336 Ile Tyr Ala Asp Gly Arg Ser Arg Ala Phe Ile Asn Gly Ala Ser Ala 100 105 110 acg gta ggc caa ttg cgc gaa atc ggc gaa acc ctc att gat att tat 384 Thr Val Gly Gln Leu Arg Glu Ile Gly Glu Thr Leu Ile Asp Ile Tyr 115 120 125 agc cag aat gcg cat cac tcc tta ctc aaa att gct acc cag cgc gag 432 Ser Gln Asn Ala His His Ser Leu Leu Lys Ile Ala Thr Gln Arg Glu 130 135 140 atc ctg gat gct tat gca caa gcg tca cca ctg gca aaa caa gtt gcc 480 Ile Leu Asp Ala Tyr Ala Gln Ala Ser Pro Leu Ala Lys Gln Val Ala 145 150 155 160 aag tta tac aac gac tgg ttt cag cta cat cag cag cag ctg gcg tat 528 Lys Leu Tyr Asn Asp Trp Phe Gln Leu His Gln Gln Gln Leu Ala Tyr 165 170 175 gag aaa aat tcc agc cag ttt gca gaa gag ctg gct gag cta cgt gac 576 Glu Lys Asn Ser Ser Gln Phe Ala Glu Glu Leu Ala Glu Leu Arg Asp 180 185 190 agt aca aga gaa ttg aag cag ttg ggt ttt gcc agt gac gaa tgg cag 624 Ser Thr Arg Glu Leu Lys Gln Leu Gly Phe Ala Ser Asp Glu Trp Gln 195 200 205 gca tta cag cag gag cat gtc cgt tta agt aac ggt gcc agt ttg ctc 672 Ala Leu Gln Gln Glu His Val Arg Leu Ser Asn Gly Ala Ser Leu Leu 210 215 220 agt ggc atg gaa gcc agc ctg caa atg atg agt gaa ggg gat gaa gtc 720 Ser Gly Met Glu Ala Ser Leu Gln Met Met Ser Glu Gly Asp Glu Val 225 230 235 240 aat gcg ctg gat ttg ctc tcc cag gca caa acc aag ctg gcc gaa ctg 768 Asn Ala Leu Asp Leu Leu Ser Gln Ala Gln Thr Lys Leu Ala Glu Leu 245 250 255 cag acg atg gat gcc gga ttg caa gcg att gct gaa aac ctt gat tca 816 Gln Thr Met Asp Ala Gly Leu Gln Ala Ile Ala Glu Asn Leu Asp Ser 260 265 270 gcc gtg gtg caa ctg gaa gaa gcc agc cgg gca ctg aat cgc tat ctg 864 Ala Val Val Gln Leu Glu Glu Ala Ser Arg Ala Leu Asn Arg Tyr Leu 275 280 285 caa aaa agt gag ctg gat cct gaa cgc ctc gct gag gtt gaa gcg cgt 912 Gln Lys Ser Glu Leu Asp Pro Glu Arg Leu Ala Glu Val Glu Ala Arg 290 295 300 atc cag gcg att cat ggc gcc gca cgc aag ttc cgc atc aag cct gat 960 Ile Gln Ala Ile His Gly Ala Ala Arg Lys Phe Arg Ile Lys Pro Asp 305 310 315 320 gag ctg cct gaa tta ttg gcc cag caa ttg cag cgg gtg gct gag ctt 1008 Glu Leu Pro Glu Leu Leu Ala Gln Gln Leu Gln Arg Val Ala Glu Leu 325 330 335 gaa ggt ttt tct gat gac ggt gcc ttg gcc aag caa gtg caa cta gcc 1056 Glu Gly Phe Ser Asp Asp Gly Ala Leu Ala Lys Gln Val Gln Leu Ala 340 345 350 tgg aaa gcc tac cat gaa caa gcc acc cag ctt tcg tca gcc agg cag 1104 Trp Lys Ala Tyr His Glu Gln Ala Thr Gln Leu Ser Ser Ala Arg Gln 355 360 365 caa gct gcg caa aga ctg gct aaa acc atc act gag caa atg cag gcg 1152 Gln Ala Ala Gln Arg Leu Ala Lys Thr Ile Thr Glu Gln Met Gln Ala 370 375 380 ttg tcc tta aag ggc ggc cag ttt gct gtg gct tta aca acc tcc agt 1200 Leu Ser Leu Lys Gly Gly Gln Phe Ala Val Ala Leu Thr Thr Ser Ser 385 390 395 400 gag ccc act gct cat gga ttg gaa caa gtg gag ttc ctg gtg gct ggt 1248 Glu Pro Thr Ala His Gly Leu Glu Gln Val Glu Phe Leu Val Ala Gly 405 410 415 cat gcc ggc gtt gag ccc cgt ccg ctc aat aaa gtt gcc tct ggt ggt 1296 His Ala Gly Val Glu Pro Arg Pro Leu Asn Lys Val Ala Ser Gly Gly 420 425 430 gag ttg tca cgc atc agc ctg gcc ttg cag gtg acc acc gct tca tta 1344 Glu Leu Ser Arg Ile Ser Leu Ala Leu Gln Val Thr Thr Ala Ser Leu 435 440 445 ggc aca gtg ccc tgc atg att ttt gat gaa gtg gat gtt ggc att ggc 1392 Gly Thr Val Pro Cys Met Ile Phe Asp Glu Val Asp Val Gly Ile Gly 450 455 460 ggc ggc gtt gca gaa gtg gtg ggc aga ttg ttg aat cag ctt ggc cag 1440 Gly Gly Val Ala Glu Val Val Gly Arg Leu Leu Asn Gln Leu Gly Gln 465 470 475 480 cat cgg cag gta ctg gtg att acc cac ctg gca caa gtt gca gca cag 1488 His Arg Gln Val Leu Val Ile Thr His Leu Ala Gln Val Ala Ala Gln 485 490 495 gcg cag cag cat ttg cag gtt agc aaa tct gag cag caa ggc gcg aca 1536 Ala Gln Gln His Leu Gln Val Ser Lys Ser Glu Gln Gln Gly Ala Thr 500 505 510 tta agc cgt att cgc gca ttg gac gca agc gag cga gta gaa gag gtg 1584 Leu Ser Arg Ile Arg Ala Leu Asp Ala Ser Glu Arg Val Glu Glu Val 515 520 525 gca cgt atg cta ggt ggt ttg cag att act gag acg aca agg aat cac 1632 Ala Arg Met Leu Gly Gly Leu Gln Ile Thr Glu Thr Thr Arg Asn His 530 535 540 gcc cgg gaa atg ctt ggg ata gtc tga 1659 Ala Arg Glu Met Leu Gly Ile Val 545 550 4 552 PRT Methylophilus methylotrophus 4 Met Leu Gln Ala Leu Ser Ile Arg Asp Phe Val Ile Val Asp Thr Leu 1 5 10 15 Glu Leu Glu Phe Ser Ala Gly Tyr Thr Ala Leu Thr Gly Glu Thr Gly 20 25 30 Ala Gly Lys Ser Ile Leu Ile Asp Ala Leu Ser Leu Ser Leu Gly Ala 35 40 45 Arg Asn Glu Gly Asp Val Thr Arg Lys Gly Cys Glu Lys Ala Glu Ile 50 55 60 Ser Thr Thr Phe Asp Ile Ala Asp Asn Leu Ala Ala Arg Asp Trp Leu 65 70 75 80 Gln Ala Gln Glu Ile Asp Ile Glu Asp Ala Leu Val Leu Arg Arg Val 85 90 95 Ile Tyr Ala Asp Gly Arg Ser Arg Ala Phe Ile Asn Gly Ala Ser Ala 100 105 110 Thr Val Gly Gln Leu Arg Glu Ile Gly Glu Thr Leu Ile Asp Ile Tyr 115 120 125 Ser Gln Asn Ala His His Ser Leu Leu Lys Ile Ala Thr Gln Arg Glu 130 135 140 Ile Leu Asp Ala Tyr Ala Gln Ala Ser Pro Leu Ala Lys Gln Val Ala 145 150 155 160 Lys Leu Tyr Asn Asp Trp Phe Gln Leu His Gln Gln Gln Leu Ala Tyr 165 170 175 Glu Lys Asn Ser Ser Gln Phe Ala Glu Glu Leu Ala Glu Leu Arg Asp 180 185 190 Ser Thr Arg Glu Leu Lys Gln Leu Gly Phe Ala Ser Asp Glu Trp Gln 195 200 205 Ala Leu Gln Gln Glu His Val Arg Leu Ser Asn Gly Ala Ser Leu Leu 210 215 220 Ser Gly Met Glu Ala Ser Leu Gln Met Met Ser Glu Gly Asp Glu Val 225 230 235 240 Asn Ala Leu Asp Leu Leu Ser Gln Ala Gln Thr Lys Leu Ala Glu Leu 245 250 255 Gln Thr Met Asp Ala Gly Leu Gln Ala Ile Ala Glu Asn Leu Asp Ser 260 265 270 Ala Val Val Gln Leu Glu Glu Ala Ser Arg Ala Leu Asn Arg Tyr Leu 275 280 285 Gln Lys Ser Glu Leu Asp Pro Glu Arg Leu Ala Glu Val Glu Ala Arg 290 295 300 Ile Gln Ala Ile His Gly Ala Ala Arg Lys Phe Arg Ile Lys Pro Asp 305 310 315 320 Glu Leu Pro Glu Leu Leu Ala Gln Gln Leu Gln Arg Val Ala Glu Leu 325 330 335 Glu Gly Phe Ser Asp Asp Gly Ala Leu Ala Lys Gln Val Gln Leu Ala 340 345 350 Trp Lys Ala Tyr His Glu Gln Ala Thr Gln Leu Ser Ser Ala Arg Gln 355 360 365 Gln Ala Ala Gln Arg Leu Ala Lys Thr Ile Thr Glu Gln Met Gln Ala 370 375 380 Leu Ser Leu Lys Gly Gly Gln Phe Ala Val Ala Leu Thr Thr Ser Ser 385 390 395 400 Glu Pro Thr Ala His Gly Leu Glu Gln Val Glu Phe Leu Val Ala Gly 405 410 415 His Ala Gly Val Glu Pro Arg Pro Leu Asn Lys Val Ala Ser Gly Gly 420 425 430 Glu Leu Ser Arg Ile Ser Leu Ala Leu Gln Val Thr Thr Ala Ser Leu 435 440 445 Gly Thr Val Pro Cys Met Ile Phe Asp Glu Val Asp Val Gly Ile Gly 450 455 460 Gly Gly Val Ala Glu Val Val Gly Arg Leu Leu Asn Gln Leu Gly Gln 465 470 475 480 His Arg Gln Val Leu Val Ile Thr His Leu Ala Gln Val Ala Ala Gln 485 490 495 Ala Gln Gln His Leu Gln Val Ser Lys Ser Glu Gln Gln Gly Ala Thr 500 505 510 Leu Ser Arg Ile Arg Ala Leu Asp Ala Ser Glu Arg Val Glu Glu Val 515 520 525 Ala Arg Met Leu Gly Gly Leu Gln Ile Thr Glu Thr Thr Arg Asn His 530 535 540 Ala Arg Glu Met Leu Gly Ile Val 545 550 5 753 DNA Methylophilus methylotrophus CDS (1)..(753) 5 atg gca cta aaa cca tct gat tca ggt agc cac aaa caa agc ctg caa 48 Met Ala Leu Lys Pro Ser Asp Ser Gly Ser His Lys Gln Ser Leu Gln 1 5 10 15 ccg gtt ttt att ctt cat acc tat ccc ttt aag gaa acc agc ctg gtc 96 Pro Val Phe Ile Leu His Thr Tyr Pro Phe Lys Glu Thr Ser Leu Val 20 25 30 gtc gaa atg ttt agt cgc gac ctg gga cgt att gcg gcc gtt gcc aag 144 Val Glu Met Phe Ser Arg Asp Leu Gly Arg Ile Ala Ala Val Ala Lys 35 40 45 ggt gca cga cgg ccc ctg tcg gcc atg cgt ggc atg ttg cag tct ttt 192 Gly Ala Arg Arg Pro Leu Ser Ala Met Arg Gly Met Leu Gln Ser Phe 50 55 60 cag cag ctg gcc ggt acc tgg tca ggc aaa aac gag ctc aaa acc ctg 240 Gln Gln Leu Ala Gly Thr Trp Ser Gly Lys Asn Glu Leu Lys Thr Leu 65 70 75 80 cat gac ctc gag tgg atg aca ggc ctg aca ttg ctc agg ggc gat gcg 288 His Asp Leu Glu Trp Met Thr Gly Leu Thr Leu Leu Arg Gly Asp Ala 85 90 95 ctc atg tgc ggt ttt tac atg aac gag ctg ttg ctg cgc ttg ctg ccg 336 Leu Met Cys Gly Phe Tyr Met Asn Glu Leu Leu Leu Arg Leu Leu Pro 100 105 110 cgg gat gat gcc cat acg cag cta ttc gag tat tac gcc caa acc att 384 Arg Asp Asp Ala His Thr Gln Leu Phe Glu Tyr Tyr Ala Gln Thr Ile 115 120 125 caa tta ttg tct acc ttg cag cag gat gcc ggc agt ggc cag ctg gcc 432 Gln Leu Leu Ser Thr Leu Gln Gln Asp Ala Gly Ser Gly Gln Leu Ala 130 135 140 gag atc atg cgc cgg ttt gaa ctc aaa atg tta cag gaa ttg ggg tac 480 Glu Ile Met Arg Arg Phe Glu Leu Lys Met Leu Gln Glu Leu Gly Tyr 145 150 155 160 gcg gta cca ctg acg cac gat gaa cat ggc gaa gtg ata cac gcc gat 528 Ala Val Pro Leu Thr His Asp Glu His Gly Glu Val Ile His Ala Asp 165 170 175 gag ata tac cgc ttt gag gcc gac tat ggc gca tgt gcc ccg tct gca 576 Glu Ile Tyr Arg Phe Glu Ala Asp Tyr Gly Ala Cys Ala Pro Ser Ala 180 185 190 acc aga aat ggg ttg ctg gtg cag ggg gca aca tta ctg gag atg gcc 624 Thr Arg Asn Gly Leu Leu Val Gln Gly Ala Thr Leu Leu Glu Met Ala 195 200 205 cgc ggc aat tac agc agc gcc acg acg caa tca caa agc aag caa ctc 672 Arg Gly Asn Tyr Ser Ser Ala Thr Thr Gln Ser Gln Ser Lys Gln Leu 210 215 220 atg cgt tac ctg cta cag cat tac ctg ggt gaa aaa ccc ttg cat acg 720 Met Arg Tyr Leu Leu Gln His Tyr Leu Gly Glu Lys Pro Leu His Thr 225 230 235 240 cgg caa ctg ctg gtg gat ttg cag gat ttc tga 753 Arg Gln Leu Leu Val Asp Leu Gln Asp Phe 245 250 6 250 PRT Methylophilus methylotrophus 6 Met Ala Leu Lys Pro Ser Asp Ser Gly Ser His Lys Gln Ser Leu Gln 1 5 10 15 Pro Val Phe Ile Leu His Thr Tyr Pro Phe Lys Glu Thr Ser Leu Val 20 25 30 Val Glu Met Phe Ser Arg Asp Leu Gly Arg Ile Ala Ala Val Ala Lys 35 40 45 Gly Ala Arg Arg Pro Leu Ser Ala Met Arg Gly Met Leu Gln Ser Phe 50 55 60 Gln Gln Leu Ala Gly Thr Trp Ser Gly Lys Asn Glu Leu Lys Thr Leu 65 70 75 80 His Asp Leu Glu Trp Met Thr Gly Leu Thr Leu Leu Arg Gly Asp

Ala 85 90 95 Leu Met Cys Gly Phe Tyr Met Asn Glu Leu Leu Leu Arg Leu Leu Pro 100 105 110 Arg Asp Asp Ala His Thr Gln Leu Phe Glu Tyr Tyr Ala Gln Thr Ile 115 120 125 Gln Leu Leu Ser Thr Leu Gln Gln Asp Ala Gly Ser Gly Gln Leu Ala 130 135 140 Glu Ile Met Arg Arg Phe Glu Leu Lys Met Leu Gln Glu Leu Gly Tyr 145 150 155 160 Ala Val Pro Leu Thr His Asp Glu His Gly Glu Val Ile His Ala Asp 165 170 175 Glu Ile Tyr Arg Phe Glu Ala Asp Tyr Gly Ala Cys Ala Pro Ser Ala 180 185 190 Thr Arg Asn Gly Leu Leu Val Gln Gly Ala Thr Leu Leu Glu Met Ala 195 200 205 Arg Gly Asn Tyr Ser Ser Ala Thr Thr Gln Ser Gln Ser Lys Gln Leu 210 215 220 Met Arg Tyr Leu Leu Gln His Tyr Leu Gly Glu Lys Pro Leu His Thr 225 230 235 240 Arg Gln Leu Leu Val Asp Leu Gln Asp Phe 245 250 7 315 DNA Methylophilus methylotrophus CDS (1)..(315) 7 atg aca ctc aca aaa gct gac ttg gcc gat ttg ttg ttt gaa caa gtc 48 Met Thr Leu Thr Lys Ala Asp Leu Ala Asp Leu Leu Phe Glu Gln Val 1 5 10 15 ggg cta aac aag cgt gaa gcc aag gac atg gta gaa gcg ttt ttt gaa 96 Gly Leu Asn Lys Arg Glu Ala Lys Asp Met Val Glu Ala Phe Phe Glu 20 25 30 gaa gtc cgt aat gca ctg gaa caa ggc gat agc gtt aaa ctg tct ggt 144 Glu Val Arg Asn Ala Leu Glu Gln Gly Asp Ser Val Lys Leu Ser Gly 35 40 45 ttt ggt aat ttt gaa ttg cgt act aaa tcc gaa cgt cct ggc cgt aac 192 Phe Gly Asn Phe Glu Leu Arg Thr Lys Ser Glu Arg Pro Gly Arg Asn 50 55 60 ccg aaa act ggc gaa gaa atc ccg atc tct gca cgt cgt gta gtg act 240 Pro Lys Thr Gly Glu Glu Ile Pro Ile Ser Ala Arg Arg Val Val Thr 65 70 75 80 ttt cac gcc agt cag aaa tta aaa ttg cgc gta gaa gag cac tac gcc 288 Phe His Ala Ser Gln Lys Leu Lys Leu Arg Val Glu Glu His Tyr Ala 85 90 95 gaa caa cct ttg caa gca caa gcc tga 315 Glu Gln Pro Leu Gln Ala Gln Ala 100 8 104 PRT Methylophilus methylotrophus 8 Met Thr Leu Thr Lys Ala Asp Leu Ala Asp Leu Leu Phe Glu Gln Val 1 5 10 15 Gly Leu Asn Lys Arg Glu Ala Lys Asp Met Val Glu Ala Phe Phe Glu 20 25 30 Glu Val Arg Asn Ala Leu Glu Gln Gly Asp Ser Val Lys Leu Ser Gly 35 40 45 Phe Gly Asn Phe Glu Leu Arg Thr Lys Ser Glu Arg Pro Gly Arg Asn 50 55 60 Pro Lys Thr Gly Glu Glu Ile Pro Ile Ser Ala Arg Arg Val Val Thr 65 70 75 80 Phe His Ala Ser Gln Lys Leu Lys Leu Arg Val Glu Glu His Tyr Ala 85 90 95 Glu Gln Pro Leu Gln Ala Gln Ala 100 9 288 DNA Methylophilus methylotrophus CDS (1)..(288) 9 atg aca cga tct gaa ctg ata gac ctg ctt gcc cag cgc ttt ccg caa 48 Met Thr Arg Ser Glu Leu Ile Asp Leu Leu Ala Gln Arg Phe Pro Gln 1 5 10 15 tta gtg ctg aaa gat gcc gag tta tcc gtc aaa acc atc ctg gat gca 96 Leu Val Leu Lys Asp Ala Glu Leu Ser Val Lys Thr Ile Leu Asp Ala 20 25 30 atg acg gaa aac ctg gcc aca ggc gag cgt atc gaa ata cgc ggt ttt 144 Met Thr Glu Asn Leu Ala Thr Gly Glu Arg Ile Glu Ile Arg Gly Phe 35 40 45 ggc agc ttc agc ctc aat tac cgc ccg ccc cgt ttg gga cgt aac ccg 192 Gly Ser Phe Ser Leu Asn Tyr Arg Pro Pro Arg Leu Gly Arg Asn Pro 50 55 60 aaa aca ggc act aag gtg caa gtt ccg gct aaa tat gtc cct cac ttt 240 Lys Thr Gly Thr Lys Val Gln Val Pro Ala Lys Tyr Val Pro His Phe 65 70 75 80 aag gct ggt aaa gag ctg cgt gac cgc gta gat gct atc gaa tct taa 288 Lys Ala Gly Lys Glu Leu Arg Asp Arg Val Asp Ala Ile Glu Ser 85 90 95 10 95 PRT Methylophilus methylotrophus 10 Met Thr Arg Ser Glu Leu Ile Asp Leu Leu Ala Gln Arg Phe Pro Gln 1 5 10 15 Leu Val Leu Lys Asp Ala Glu Leu Ser Val Lys Thr Ile Leu Asp Ala 20 25 30 Met Thr Glu Asn Leu Ala Thr Gly Glu Arg Ile Glu Ile Arg Gly Phe 35 40 45 Gly Ser Phe Ser Leu Asn Tyr Arg Pro Pro Arg Leu Gly Arg Asn Pro 50 55 60 Lys Thr Gly Thr Lys Val Gln Val Pro Ala Lys Tyr Val Pro His Phe 65 70 75 80 Lys Ala Gly Lys Glu Leu Arg Asp Arg Val Asp Ala Ile Glu Ser 85 90 95 11 1950 DNA Methylophilus methylotrophus CDS (1)..(1950) 11 atg gcc aga ccc aaa gca gat aaa agt tta gat aaa gaa ctc gga atc 48 Met Ala Arg Pro Lys Ala Asp Lys Ser Leu Asp Lys Glu Leu Gly Ile 1 5 10 15 aac gaa gtc gaa acc cct aaa gat ccc gct acc cag gca ctg gat gcc 96 Asn Glu Val Glu Thr Pro Lys Asp Pro Ala Thr Gln Ala Leu Asp Ala 20 25 30 gag gca cgc cgt acc cgc ctg aaa acc ctg att gtc ctg ggt aaa gag 144 Glu Ala Arg Arg Thr Arg Leu Lys Thr Leu Ile Val Leu Gly Lys Glu 35 40 45 cgt ggc tac ctg acc tac gcc gaa atc aat gac cac ctg ccg gac gat 192 Arg Gly Tyr Leu Thr Tyr Ala Glu Ile Asn Asp His Leu Pro Asp Asp 50 55 60 gtg caa gac tcc gag cag atc gag agc att atc ggc atg atc aat gac 240 Val Gln Asp Ser Glu Gln Ile Glu Ser Ile Ile Gly Met Ile Asn Asp 65 70 75 80 atg ggt atc cag gtc tat gaa gaa gcg ccg gat gcc gaa gtg ttg ttg 288 Met Gly Ile Gln Val Tyr Glu Glu Ala Pro Asp Ala Glu Val Leu Leu 85 90 95 atg tct gat gcg cca cca gct gtg act gat gat gat gcg gca gaa gag 336 Met Ser Asp Ala Pro Pro Ala Val Thr Asp Asp Asp Ala Ala Glu Glu 100 105 110 gct gag cag gcc ttg gca acc gtg gac tct gaa ttt ggt cgc acg act 384 Ala Glu Gln Ala Leu Ala Thr Val Asp Ser Glu Phe Gly Arg Thr Thr 115 120 125 gac cca gta cgt atg tac atg cgc gaa atg ggg act gtt gac ctg ttg 432 Asp Pro Val Arg Met Tyr Met Arg Glu Met Gly Thr Val Asp Leu Leu 130 135 140 acg cgt gag ggc gaa atc gag atc gcc aaa cgc att gag gat ggc ctc 480 Thr Arg Glu Gly Glu Ile Glu Ile Ala Lys Arg Ile Glu Asp Gly Leu 145 150 155 160 aag cac atg gtg cag gcg att gca gcc tgt cca acg acc atc gcc cag 528 Lys His Met Val Gln Ala Ile Ala Ala Cys Pro Thr Thr Ile Ala Gln 165 170 175 ttg ctg gag atg gtc gac aag gtt gaa aaa gac gaa atg agc gtt gat 576 Leu Leu Glu Met Val Asp Lys Val Glu Lys Asp Glu Met Ser Val Asp 180 185 190 gaa ctg gta gac ggt ttg atc gat agt gat ttg ggt ctg gat gac gcg 624 Glu Leu Val Asp Gly Leu Ile Asp Ser Asp Leu Gly Leu Asp Asp Ala 195 200 205 ctg gcg gcc gca gat gtt gag gaa gaa gaa gcc gac gac gaa gaa gat 672 Leu Ala Ala Ala Asp Val Glu Glu Glu Glu Ala Asp Asp Glu Glu Asp 210 215 220 gaa gat gat gaa gaa gat ggt gat gcc aaa gcg tct gcc atg tct gcc 720 Glu Asp Asp Glu Glu Asp Gly Asp Ala Lys Ala Ser Ala Met Ser Ala 225 230 235 240 gaa gcg ctg gcc aaa ctc aag gat gaa gtc ttg tca cgc ttt gcc gtc 768 Glu Ala Leu Ala Lys Leu Lys Asp Glu Val Leu Ser Arg Phe Ala Val 245 250 255 atc cgt gct gcc aac gca aaa atg aac acc atc cgt gag gat aaa ggc 816 Ile Arg Ala Ala Asn Ala Lys Met Asn Thr Ile Arg Glu Asp Lys Gly 260 265 270 acc agc gat gca gaa tat aaa act ttg cat cag caa gta ctg gat gag 864 Thr Ser Asp Ala Glu Tyr Lys Thr Leu His Gln Gln Val Leu Asp Glu 275 280 285 ctg acc gcc ttc cgt ttc tct gcc aaa cag gtt gag gcc ttg tgc gac 912 Leu Thr Ala Phe Arg Phe Ser Ala Lys Gln Val Glu Ala Leu Cys Asp 290 295 300 cag gtg cgt aac atg gtg gaa gag gtg cgt acg cac gaa cgc aag atc 960 Gln Val Arg Asn Met Val Glu Glu Val Arg Thr His Glu Arg Lys Ile 305 310 315 320 atg gag ttt tgt gta gac aag gcc aat atg ccg cgt cag caa ttc atc 1008 Met Glu Phe Cys Val Asp Lys Ala Asn Met Pro Arg Gln Gln Phe Ile 325 330 335 aag agc ttt gtc ggt aac gaa atc aac ctt aac tgg ctc gat gac gag 1056 Lys Ser Phe Val Gly Asn Glu Ile Asn Leu Asn Trp Leu Asp Asp Glu 340 345 350 ctg gca ggc aaa cct gcc tat ggt gag cgt tta gaa cgt ttc agg cac 1104 Leu Ala Gly Lys Pro Ala Tyr Gly Glu Arg Leu Glu Arg Phe Arg His 355 360 365 tct atc cta gac cag caa gag cgt tta caa aac ctg cag aac cgc gtg 1152 Ser Ile Leu Asp Gln Gln Glu Arg Leu Gln Asn Leu Gln Asn Arg Val 370 375 380 gga ttg cca atc aaa gag ctg cgt gaa atc aac aag caa atg acc act 1200 Gly Leu Pro Ile Lys Glu Leu Arg Glu Ile Asn Lys Gln Met Thr Thr 385 390 395 400 ggc gaa gct cgt gca cga cgc gcc aag cgc gaa atg atc gag gcc aac 1248 Gly Glu Ala Arg Ala Arg Arg Ala Lys Arg Glu Met Ile Glu Ala Asn 405 410 415 ctg cgt ctg gtg atc tcc att gca aaa aaa tac acc aat cgt ggt ttg 1296 Leu Arg Leu Val Ile Ser Ile Ala Lys Lys Tyr Thr Asn Arg Gly Leu 420 425 430 cag ttc ctc gac ttg atc caa gag ggc aac atc ggc ttg atg aaa gcc 1344 Gln Phe Leu Asp Leu Ile Gln Glu Gly Asn Ile Gly Leu Met Lys Ala 435 440 445 gtg gat aag ttt gaa tac cgc cgt ggt tac aaa ttt tct acc tat gcg 1392 Val Asp Lys Phe Glu Tyr Arg Arg Gly Tyr Lys Phe Ser Thr Tyr Ala 450 455 460 aca tgg tgg att cgc cag gcg att acc cgt tca atc gca gac cag gca 1440 Thr Trp Trp Ile Arg Gln Ala Ile Thr Arg Ser Ile Ala Asp Gln Ala 465 470 475 480 cgt acc atc cgt atc ccg gtc cac atg atc gaa acc atc aac aag atg 1488 Arg Thr Ile Arg Ile Pro Val His Met Ile Glu Thr Ile Asn Lys Met 485 490 495 aac cgt atc agc cgc cag atc ttg caa gaa act ggc ctg gaa cct gat 1536 Asn Arg Ile Ser Arg Gln Ile Leu Gln Glu Thr Gly Leu Glu Pro Asp 500 505 510 ccg gct act ctg gct gaa aaa atg gat atg ccg gaa gaa aaa atc cgc 1584 Pro Ala Thr Leu Ala Glu Lys Met Asp Met Pro Glu Glu Lys Ile Arg 515 520 525 aaa atc ctc aaa atc agc aaa gag ccg atc tca atg gaa acg ccg att 1632 Lys Ile Leu Lys Ile Ser Lys Glu Pro Ile Ser Met Glu Thr Pro Ile 530 535 540 ggt gac gat gaa gac agc cac ttg ggc gac ttt att gaa gac aac acc 1680 Gly Asp Asp Glu Asp Ser His Leu Gly Asp Phe Ile Glu Asp Asn Thr 545 550 555 560 aca ctg gca ccg atg gac gca gct gtt tac gcc agc ctg cgt gac gcc 1728 Thr Leu Ala Pro Met Asp Ala Ala Val Tyr Ala Ser Leu Arg Asp Ala 565 570 575 acc agt gaa gtg ctg gaa tca ctg acc cca cgt gaa gcc aaa gtg ctg 1776 Thr Ser Glu Val Leu Glu Ser Leu Thr Pro Arg Glu Ala Lys Val Leu 580 585 590 cgc atg cgc ttt ggt atc gaa atg aac acc gac cac acg ctg gaa gaa 1824 Arg Met Arg Phe Gly Ile Glu Met Asn Thr Asp His Thr Leu Glu Glu 595 600 605 gtt ggc aaa caa ttt gac gtc acc cgt gaa cgt atc cgc cag att gaa 1872 Val Gly Lys Gln Phe Asp Val Thr Arg Glu Arg Ile Arg Gln Ile Glu 610 615 620 gct aag gcc ctg cgt aaa ctg cgt cac ccg acc cgc tcg gaa cgc ctg 1920 Ala Lys Ala Leu Arg Lys Leu Arg His Pro Thr Arg Ser Glu Arg Leu 625 630 635 640 cgc agc ttc tta gaa aac ggt cag gat tag 1950 Arg Ser Phe Leu Glu Asn Gly Gln Asp 645 12 649 PRT Methylophilus methylotrophus 12 Met Ala Arg Pro Lys Ala Asp Lys Ser Leu Asp Lys Glu Leu Gly Ile 1 5 10 15 Asn Glu Val Glu Thr Pro Lys Asp Pro Ala Thr Gln Ala Leu Asp Ala 20 25 30 Glu Ala Arg Arg Thr Arg Leu Lys Thr Leu Ile Val Leu Gly Lys Glu 35 40 45 Arg Gly Tyr Leu Thr Tyr Ala Glu Ile Asn Asp His Leu Pro Asp Asp 50 55 60 Val Gln Asp Ser Glu Gln Ile Glu Ser Ile Ile Gly Met Ile Asn Asp 65 70 75 80 Met Gly Ile Gln Val Tyr Glu Glu Ala Pro Asp Ala Glu Val Leu Leu 85 90 95 Met Ser Asp Ala Pro Pro Ala Val Thr Asp Asp Asp Ala Ala Glu Glu 100 105 110 Ala Glu Gln Ala Leu Ala Thr Val Asp Ser Glu Phe Gly Arg Thr Thr 115 120 125 Asp Pro Val Arg Met Tyr Met Arg Glu Met Gly Thr Val Asp Leu Leu 130 135 140 Thr Arg Glu Gly Glu Ile Glu Ile Ala Lys Arg Ile Glu Asp Gly Leu 145 150 155 160 Lys His Met Val Gln Ala Ile Ala Ala Cys Pro Thr Thr Ile Ala Gln 165 170 175 Leu Leu Glu Met Val Asp Lys Val Glu Lys Asp Glu Met Ser Val Asp 180 185 190 Glu Leu Val Asp Gly Leu Ile Asp Ser Asp Leu Gly Leu Asp Asp Ala 195 200 205 Leu Ala Ala Ala Asp Val Glu Glu Glu Glu Ala Asp Asp Glu Glu Asp 210 215 220 Glu Asp Asp Glu Glu Asp Gly Asp Ala Lys Ala Ser Ala Met Ser Ala 225 230 235 240 Glu Ala Leu Ala Lys Leu Lys Asp Glu Val Leu Ser Arg Phe Ala Val 245 250 255 Ile Arg Ala Ala Asn Ala Lys Met Asn Thr Ile Arg Glu Asp Lys Gly 260 265 270 Thr Ser Asp Ala Glu Tyr Lys Thr Leu His Gln Gln Val Leu Asp Glu 275 280 285 Leu Thr Ala Phe Arg Phe Ser Ala Lys Gln Val Glu Ala Leu Cys Asp 290 295 300 Gln Val Arg Asn Met Val Glu Glu Val Arg Thr His Glu Arg Lys Ile 305 310 315 320 Met Glu Phe Cys Val Asp Lys Ala Asn Met Pro Arg Gln Gln Phe Ile 325 330 335 Lys Ser Phe Val Gly Asn Glu Ile Asn Leu Asn Trp Leu Asp Asp Glu 340 345 350 Leu Ala Gly Lys Pro Ala Tyr Gly Glu Arg Leu Glu Arg Phe Arg His 355 360 365 Ser Ile Leu Asp Gln Gln Glu Arg Leu Gln Asn Leu Gln Asn Arg Val 370 375 380 Gly Leu Pro Ile Lys Glu Leu Arg Glu Ile Asn Lys Gln Met Thr Thr 385 390 395 400 Gly Glu Ala Arg Ala Arg Arg Ala Lys Arg Glu Met Ile Glu Ala Asn 405 410 415 Leu Arg Leu Val Ile Ser Ile Ala Lys Lys Tyr Thr Asn Arg Gly Leu 420 425 430 Gln Phe Leu Asp Leu Ile Gln Glu Gly Asn Ile Gly Leu Met Lys Ala 435 440 445 Val Asp Lys Phe Glu Tyr Arg Arg Gly Tyr Lys Phe Ser Thr Tyr Ala 450 455 460 Thr Trp Trp Ile Arg Gln Ala Ile Thr Arg Ser Ile Ala Asp Gln Ala 465 470 475 480 Arg Thr Ile Arg Ile Pro Val His Met Ile Glu Thr Ile Asn Lys Met 485 490 495 Asn Arg Ile Ser Arg Gln Ile Leu Gln Glu Thr Gly Leu Glu Pro Asp 500 505 510 Pro Ala Thr Leu Ala Glu Lys Met Asp Met Pro Glu Glu Lys Ile Arg 515 520 525 Lys Ile Leu Lys Ile Ser Lys Glu Pro Ile Ser Met Glu Thr Pro Ile 530 535 540 Gly Asp Asp Glu Asp Ser His Leu Gly Asp Phe Ile Glu Asp Asn Thr 545 550 555 560 Thr Leu Ala Pro Met Asp Ala Ala Val Tyr Ala Ser Leu Arg Asp Ala 565 570 575 Thr Ser Glu Val Leu Glu Ser Leu Thr Pro Arg Glu Ala Lys Val Leu 580 585 590 Arg Met Arg Phe Gly Ile Glu Met Asn Thr Asp His Thr Leu Glu Glu 595 600 605 Val Gly Lys Gln Phe Asp Val Thr Arg Glu Arg Ile Arg Gln Ile Glu 610 615 620 Ala Lys Ala Leu Arg Lys Leu Arg His Pro Thr Arg Ser Glu Arg Leu 625 630 635 640 Arg Ser Phe Leu Glu Asn Gly Gln Asp 645 13 741 DNA Methylophilus methylotrophus CDS (1)..(741) 13 atg tta cac aca gca aat tta gaa gcg tta aca cgc acc ggg ttc gtc 48 Met Leu His Thr Ala Asn Leu Glu Ala Leu Thr Arg Thr Gly Phe Val 1 5

10 15 gaa aat aac aca aaa gcg tat act act ccc aat cat aaa att acc caa 96 Glu Asn Asn Thr Lys Ala Tyr Thr Thr Pro Asn His Lys Ile Thr Gln 20 25 30 ggg agt cat cgc gat atg act gtt cag gcg gca agc gcc aaa cag caa 144 Gly Ser His Arg Asp Met Thr Val Gln Ala Ala Ser Ala Lys Gln Gln 35 40 45 gag aac aaa gag ttt gac cag atg ctg gtc gag cgt gct cag cag ggc 192 Glu Asn Lys Glu Phe Asp Gln Met Leu Val Glu Arg Ala Gln Gln Gly 50 55 60 gac aag cgt gcc ttt gga ttg ttg gtg gaa aaa tat cac cgt aag ctt 240 Asp Lys Arg Ala Phe Gly Leu Leu Val Glu Lys Tyr His Arg Lys Leu 65 70 75 80 gga cgc ctt tta tcg cgc atg att cgc gat cag gcg gaa gtg gaa gat 288 Gly Arg Leu Leu Ser Arg Met Ile Arg Asp Gln Ala Glu Val Glu Asp 85 90 95 gtg gtg cag gag tct ttt atc aag gct tat cgc gcg ttg cac agc ttc 336 Val Val Gln Glu Ser Phe Ile Lys Ala Tyr Arg Ala Leu His Ser Phe 100 105 110 cgt ggc gac agt gca ttt tat aca tgg ttg tat cgt att ggc att aat 384 Arg Gly Asp Ser Ala Phe Tyr Thr Trp Leu Tyr Arg Ile Gly Ile Asn 115 120 125 act gcc aaa aac tac ctg gtc tcc atg ggc cgc aag cca caa gtg ctg 432 Thr Ala Lys Asn Tyr Leu Val Ser Met Gly Arg Lys Pro Gln Val Leu 130 135 140 caa gac gtg gaa atc gaa gat gta gaa aac ttc gat gaa ggc gat gat 480 Gln Asp Val Glu Ile Glu Asp Val Glu Asn Phe Asp Glu Gly Asp Asp 145 150 155 160 atg cgc aca ctg gag acc cca gaa acg tcc ctg atg acg aaa gag att 528 Met Arg Thr Leu Glu Thr Pro Glu Thr Ser Leu Met Thr Lys Glu Ile 165 170 175 gcg caa acg gtc aat gat gcg att gcg gcg ttg cct gaa gag ttg cgt 576 Ala Gln Thr Val Asn Asp Ala Ile Ala Ala Leu Pro Glu Glu Leu Arg 180 185 190 act gca att aca ttg cgc gag ctt gaa gga ttg agt tat gaa gac att 624 Thr Ala Ile Thr Leu Arg Glu Leu Glu Gly Leu Ser Tyr Glu Asp Ile 195 200 205 gct aac gtg atg caa tgt ccg atc ggg act gtg cgt tca cgt att ttc 672 Ala Asn Val Met Gln Cys Pro Ile Gly Thr Val Arg Ser Arg Ile Phe 210 215 220 cgg gcg cgt gaa acg att gcg acc aaa ttg aga ccg ttg ttg gat acg 720 Arg Ala Arg Glu Thr Ile Ala Thr Lys Leu Arg Pro Leu Leu Asp Thr 225 230 235 240 cca cag cac aag cgc tgg tag 741 Pro Gln His Lys Arg Trp 245 14 246 PRT Methylophilus methylotrophus 14 Met Leu His Thr Ala Asn Leu Glu Ala Leu Thr Arg Thr Gly Phe Val 1 5 10 15 Glu Asn Asn Thr Lys Ala Tyr Thr Thr Pro Asn His Lys Ile Thr Gln 20 25 30 Gly Ser His Arg Asp Met Thr Val Gln Ala Ala Ser Ala Lys Gln Gln 35 40 45 Glu Asn Lys Glu Phe Asp Gln Met Leu Val Glu Arg Ala Gln Gln Gly 50 55 60 Asp Lys Arg Ala Phe Gly Leu Leu Val Glu Lys Tyr His Arg Lys Leu 65 70 75 80 Gly Arg Leu Leu Ser Arg Met Ile Arg Asp Gln Ala Glu Val Glu Asp 85 90 95 Val Val Gln Glu Ser Phe Ile Lys Ala Tyr Arg Ala Leu His Ser Phe 100 105 110 Arg Gly Asp Ser Ala Phe Tyr Thr Trp Leu Tyr Arg Ile Gly Ile Asn 115 120 125 Thr Ala Lys Asn Tyr Leu Val Ser Met Gly Arg Lys Pro Gln Val Leu 130 135 140 Gln Asp Val Glu Ile Glu Asp Val Glu Asn Phe Asp Glu Gly Asp Asp 145 150 155 160 Met Arg Thr Leu Glu Thr Pro Glu Thr Ser Leu Met Thr Lys Glu Ile 165 170 175 Ala Gln Thr Val Asn Asp Ala Ile Ala Ala Leu Pro Glu Glu Leu Arg 180 185 190 Thr Ala Ile Thr Leu Arg Glu Leu Glu Gly Leu Ser Tyr Glu Asp Ile 195 200 205 Ala Asn Val Met Gln Cys Pro Ile Gly Thr Val Arg Ser Arg Ile Phe 210 215 220 Arg Ala Arg Glu Thr Ile Ala Thr Lys Leu Arg Pro Leu Leu Asp Thr 225 230 235 240 Pro Gln His Lys Arg Trp 245 15 777 DNA Methylophilus methylotrophus CDS (1)..(777) 15 gtg ctc acg gca gag gaa gag tat ggt tac gct acc cgc ctg aaa gaa 48 Val Leu Thr Ala Glu Glu Glu Tyr Gly Tyr Ala Thr Arg Leu Lys Glu 1 5 10 15 agc ggt gac ctg gaa tct gcg cgc gca ctg att gtg tcg cat ttg cgt 96 Ser Gly Asp Leu Glu Ser Ala Arg Ala Leu Ile Val Ser His Leu Arg 20 25 30 ctg gtc gcc agt att gcg cgt ggt tac agc ggt tat ggc ttg cca cag 144 Leu Val Ala Ser Ile Ala Arg Gly Tyr Ser Gly Tyr Gly Leu Pro Gln 35 40 45 tcc gac ctg att cag gaa ggc aat att ggc ctg atg aaa gcg gtt aaa 192 Ser Asp Leu Ile Gln Glu Gly Asn Ile Gly Leu Met Lys Ala Val Lys 50 55 60 cgc ttt gac ccg gat cgt ggc gtg cgc ctg gtt tca ttt gcc atg cac 240 Arg Phe Asp Pro Asp Arg Gly Val Arg Leu Val Ser Phe Ala Met His 65 70 75 80 tgg atc aaa gcc gag att cac gaa tac att gtg cgc aac tgg cgc ctg 288 Trp Ile Lys Ala Glu Ile His Glu Tyr Ile Val Arg Asn Trp Arg Leu 85 90 95 gtg aaa aca gcc acg acc aag gca cag cgc aaa ctg ttc ttc aac ctg 336 Val Lys Thr Ala Thr Thr Lys Ala Gln Arg Lys Leu Phe Phe Asn Leu 100 105 110 cgt agc atg aag caa ggg ttt gaa acc ttg agc cag gat gat gtg cat 384 Arg Ser Met Lys Gln Gly Phe Glu Thr Leu Ser Gln Asp Asp Val His 115 120 125 cgc atc gcg acc gag ctc aat gtg aaa ccg gaa gaa gtg act gaa atg 432 Arg Ile Ala Thr Glu Leu Asn Val Lys Pro Glu Glu Val Thr Glu Met 130 135 140 gaa tat cgc ctc aat ggt cag gag att tca ctg gat gcg cag gta gat 480 Glu Tyr Arg Leu Asn Gly Gln Glu Ile Ser Leu Asp Ala Gln Val Asp 145 150 155 160 gaa gat ggc gaa gaa gta tat agc cct att tcc ttc ctg gag gat gaa 528 Glu Asp Gly Glu Glu Val Tyr Ser Pro Ile Ser Phe Leu Glu Asp Glu 165 170 175 ggc cct gag cct tct acg ctg ctg gaa aat ctg cag aat gag cac atg 576 Gly Pro Glu Pro Ser Thr Leu Leu Glu Asn Leu Gln Asn Glu His Met 180 185 190 cag act gaa ggc tta agc aac gca ttg cag caa ctt gat gag cgt agc 624 Gln Thr Glu Gly Leu Ser Asn Ala Leu Gln Gln Leu Asp Glu Arg Ser 195 200 205 cgc cgc gta ttg cag gca cgc tgg ctg acc gaa ggt gat tca gcc acc 672 Arg Arg Val Leu Gln Ala Arg Trp Leu Thr Glu Gly Asp Ser Ala Thr 210 215 220 ttg cat gag ctg gct gca gaa ttc aat gtg tcg gct gag cgt atc cgc 720 Leu His Glu Leu Ala Ala Glu Phe Asn Val Ser Ala Glu Arg Ile Arg 225 230 235 240 cag att gag caa aaa gcg atg caa aaa att aaa acc tat atg ctg gaa 768 Gln Ile Glu Gln Lys Ala Met Gln Lys Ile Lys Thr Tyr Met Leu Glu 245 250 255 agc aaa taa 777 Ser Lys 16 258 PRT Methylophilus methylotrophus 16 Val Leu Thr Ala Glu Glu Glu Tyr Gly Tyr Ala Thr Arg Leu Lys Glu 1 5 10 15 Ser Gly Asp Leu Glu Ser Ala Arg Ala Leu Ile Val Ser His Leu Arg 20 25 30 Leu Val Ala Ser Ile Ala Arg Gly Tyr Ser Gly Tyr Gly Leu Pro Gln 35 40 45 Ser Asp Leu Ile Gln Glu Gly Asn Ile Gly Leu Met Lys Ala Val Lys 50 55 60 Arg Phe Asp Pro Asp Arg Gly Val Arg Leu Val Ser Phe Ala Met His 65 70 75 80 Trp Ile Lys Ala Glu Ile His Glu Tyr Ile Val Arg Asn Trp Arg Leu 85 90 95 Val Lys Thr Ala Thr Thr Lys Ala Gln Arg Lys Leu Phe Phe Asn Leu 100 105 110 Arg Ser Met Lys Gln Gly Phe Glu Thr Leu Ser Gln Asp Asp Val His 115 120 125 Arg Ile Ala Thr Glu Leu Asn Val Lys Pro Glu Glu Val Thr Glu Met 130 135 140 Glu Tyr Arg Leu Asn Gly Gln Glu Ile Ser Leu Asp Ala Gln Val Asp 145 150 155 160 Glu Asp Gly Glu Glu Val Tyr Ser Pro Ile Ser Phe Leu Glu Asp Glu 165 170 175 Gly Pro Glu Pro Ser Thr Leu Leu Glu Asn Leu Gln Asn Glu His Met 180 185 190 Gln Thr Glu Gly Leu Ser Asn Ala Leu Gln Gln Leu Asp Glu Arg Ser 195 200 205 Arg Arg Val Leu Gln Ala Arg Trp Leu Thr Glu Gly Asp Ser Ala Thr 210 215 220 Leu His Glu Leu Ala Ala Glu Phe Asn Val Ser Ala Glu Arg Ile Arg 225 230 235 240 Gln Ile Glu Gln Lys Ala Met Gln Lys Ile Lys Thr Tyr Met Leu Glu 245 250 255 Ser Lys 17 1551 DNA Methylophilus methylotrophus CDS (1)..(1551) 17 atg aag caa aac ctg caa tta cgc att tca caa aac ctg gca ctg act 48 Met Lys Gln Asn Leu Gln Leu Arg Ile Ser Gln Asn Leu Ala Leu Thr 1 5 10 15 ccg cag cta caa cag tct atc cgt ctg tta cag tta tcc aca ttg gag 96 Pro Gln Leu Gln Gln Ser Ile Arg Leu Leu Gln Leu Ser Thr Leu Glu 20 25 30 ctc agc cag gag ctg gaa acc att ctt cag gaa aac ccg ttg ctg gag 144 Leu Ser Gln Glu Leu Glu Thr Ile Leu Gln Glu Asn Pro Leu Leu Glu 35 40 45 atg gcc gat gga gaa gaa ggc gag ttt gag gac aat tca gca acc cct 192 Met Ala Asp Gly Glu Glu Gly Glu Phe Glu Asp Asn Ser Ala Thr Pro 50 55 60 acc gaa act ata gaa tca gtc cat gca gac gat gcc aac tcg ttt gac 240 Thr Glu Thr Ile Glu Ser Val His Ala Asp Asp Ala Asn Ser Phe Asp 65 70 75 80 ctg gcc acc cag caa gaa att act gca cct gcc gag acg ctg cgc gaa 288 Leu Ala Thr Gln Gln Glu Ile Thr Ala Pro Ala Glu Thr Leu Arg Glu 85 90 95 gac tta cat gat gaa ctt ggc agc aat gaa ggc gaa ctg gct aat ctt 336 Asp Leu His Asp Glu Leu Gly Ser Asn Glu Gly Glu Leu Ala Asn Leu 100 105 110 agc gaa gaa ttc aat cct ccc gaa ttt gaa gat aat tac gag gag ttt 384 Ser Glu Glu Phe Asn Pro Pro Glu Phe Glu Asp Asn Tyr Glu Glu Phe 115 120 125 ggc agc acc agc aac tgg gat gag gca ggc cgc aac aac ctt gat gat 432 Gly Ser Thr Ser Asn Trp Asp Glu Ala Gly Arg Asn Asn Leu Asp Asp 130 135 140 gag gat agt gat ttt tcg cgt cag gat gcc agc aat atc agc ctg cgc 480 Glu Asp Ser Asp Phe Ser Arg Gln Asp Ala Ser Asn Ile Ser Leu Arg 145 150 155 160 gag cac ttg ctg gat cag att caa ctg gcg cat ttg tca cag cgg gac 528 Glu His Leu Leu Asp Gln Ile Gln Leu Ala His Leu Ser Gln Arg Asp 165 170 175 atg acg ctg gtc aag tta ctg ctc gac agc att aat gac gac ggc tac 576 Met Thr Leu Val Lys Leu Leu Leu Asp Ser Ile Asn Asp Asp Gly Tyr 180 185 190 ctt gag caa gac ttg cag gaa att gtt gaa cac ctg ccg att gag ctc 624 Leu Glu Gln Asp Leu Gln Glu Ile Val Glu His Leu Pro Ile Glu Leu 195 200 205 gaa gtc gag ctg cta gaa ctt gag acc gca ctc aaa ctg ata cag aat 672 Glu Val Glu Leu Leu Glu Leu Glu Thr Ala Leu Lys Leu Ile Gln Asn 210 215 220 ctt gat ccg gta ggt gtc ggc gcc cgt gac ttg cgg gaa tgc ctg tta 720 Leu Asp Pro Val Gly Val Gly Ala Arg Asp Leu Arg Glu Cys Leu Leu 225 230 235 240 ttg caa ctg caa cac ttg ccg gca gag acg cct tat tta cgt acg gcc 768 Leu Gln Leu Gln His Leu Pro Ala Glu Thr Pro Tyr Leu Arg Thr Ala 245 250 255 atg gca ctc gca aaa gat cac ttg gcc ctc ctg gcc aac aaa gac ttt 816 Met Ala Leu Ala Lys Asp His Leu Ala Leu Leu Ala Asn Lys Asp Phe 260 265 270 gtc aaa ctg cgc aaa ctg ctc agt tgc gat gaa acc gca ctt aaa ggt 864 Val Lys Leu Arg Lys Leu Leu Ser Cys Asp Glu Thr Ala Leu Lys Gly 275 280 285 gcg caa caa ctg ata cgc cag caa aac ccc aaa cct ggc agc gaa ttt 912 Ala Gln Gln Leu Ile Arg Gln Gln Asn Pro Lys Pro Gly Ser Glu Phe 290 295 300 gcc acc ttt agt cac gac cac ttt atc cag cat gat gtg gtg gtc aaa 960 Ala Thr Phe Ser His Asp His Phe Ile Gln His Asp Val Val Val Lys 305 310 315 320 aaa atc aaa ggt atc tgg gtc gca tcg ctc aac gat ggc gtg att ccc 1008 Lys Ile Lys Gly Ile Trp Val Ala Ser Leu Asn Asp Gly Val Ile Pro 325 330 335 aaa ctg cgg atc aac cag ctg tat gct gac atc ctc aaa cgc aac cgc 1056 Lys Leu Arg Ile Asn Gln Leu Tyr Ala Asp Ile Leu Lys Arg Asn Arg 340 345 350 gaa agc tca ggc cag tat ctg caa agc cag atg cag gaa gcg aaa tgg 1104 Glu Ser Ser Gly Gln Tyr Leu Gln Ser Gln Met Gln Glu Ala Lys Trp 355 360 365 atg atc aaa aat atc cag cag cgc ttc tcc acc att ctg cgc gtg tcg 1152 Met Ile Lys Asn Ile Gln Gln Arg Phe Ser Thr Ile Leu Arg Val Ser 370 375 380 caa gcg att gta gac cgt cag cgc aat ttc ttt gag cat ggc gac att 1200 Gln Ala Ile Val Asp Arg Gln Arg Asn Phe Phe Glu His Gly Asp Ile 385 390 395 400 gcc atg cgg cca ttg gtg ctt cgt gaa att gcc gaa gag cta gat ttg 1248 Ala Met Arg Pro Leu Val Leu Arg Glu Ile Ala Glu Glu Leu Asp Leu 405 410 415 cat gaa agt act gtt tca cgg gtc acc acg cat aaa tat atg ctc aca 1296 His Glu Ser Thr Val Ser Arg Val Thr Thr His Lys Tyr Met Leu Thr 420 425 430 cca cgt ggc gtt tac gag ctc aaa tat ttc ttt ggc agt tcg gtg gcg 1344 Pro Arg Gly Val Tyr Glu Leu Lys Tyr Phe Phe Gly Ser Ser Val Ala 435 440 445 act gat gcg ggg ggc tct tgc tca gcg acg gcc atc cgc gcc ttg atc 1392 Thr Asp Ala Gly Gly Ser Cys Ser Ala Thr Ala Ile Arg Ala Leu Ile 450 455 460 aag caa atg gtg gcc gaa gaa aat ccg aaa aaa ccg ctt tcg gat aac 1440 Lys Gln Met Val Ala Glu Glu Asn Pro Lys Lys Pro Leu Ser Asp Asn 465 470 475 480 cag atc aca gat aca ttg gct caa caa ggc att gtg gtc gcg cgc cgc 1488 Gln Ile Thr Asp Thr Leu Ala Gln Gln Gly Ile Val Val Ala Arg Arg 485 490 495 acc att gcc aaa tac cgt gag tct tta aat att ccg ccc gcc aat tta 1536 Thr Ile Ala Lys Tyr Arg Glu Ser Leu Asn Ile Pro Pro Ala Asn Leu 500 505 510 cgc aaa tcg ctt taa 1551 Arg Lys Ser Leu 515 18 516 PRT Methylophilus methylotrophus 18 Met Lys Gln Asn Leu Gln Leu Arg Ile Ser Gln Asn Leu Ala Leu Thr 1 5 10 15 Pro Gln Leu Gln Gln Ser Ile Arg Leu Leu Gln Leu Ser Thr Leu Glu 20 25 30 Leu Ser Gln Glu Leu Glu Thr Ile Leu Gln Glu Asn Pro Leu Leu Glu 35 40 45 Met Ala Asp Gly Glu Glu Gly Glu Phe Glu Asp Asn Ser Ala Thr Pro 50 55 60 Thr Glu Thr Ile Glu Ser Val His Ala Asp Asp Ala Asn Ser Phe Asp 65 70 75 80 Leu Ala Thr Gln Gln Glu Ile Thr Ala Pro Ala Glu Thr Leu Arg Glu 85 90 95 Asp Leu His Asp Glu Leu Gly Ser Asn Glu Gly Glu Leu Ala Asn Leu 100 105 110 Ser Glu Glu Phe Asn Pro Pro Glu Phe Glu Asp Asn Tyr Glu Glu Phe 115 120 125 Gly Ser Thr Ser Asn Trp Asp Glu Ala Gly Arg Asn Asn Leu Asp Asp 130 135 140 Glu Asp Ser Asp Phe Ser Arg Gln Asp Ala Ser Asn Ile Ser Leu Arg 145 150 155 160 Glu His Leu Leu Asp Gln Ile Gln Leu Ala His Leu Ser Gln Arg Asp 165 170 175 Met Thr Leu Val Lys Leu Leu Leu Asp Ser Ile Asn Asp Asp Gly Tyr 180 185 190 Leu Glu Gln Asp Leu Gln Glu Ile Val Glu His Leu Pro Ile Glu Leu 195 200 205 Glu Val Glu Leu Leu Glu Leu Glu Thr Ala Leu Lys Leu Ile Gln Asn 210 215 220 Leu Asp Pro Val Gly Val Gly Ala Arg Asp Leu Arg Glu Cys Leu Leu 225 230 235 240 Leu Gln Leu Gln His Leu Pro Ala Glu Thr Pro Tyr Leu Arg Thr Ala 245 250 255 Met Ala Leu Ala Lys Asp His Leu Ala Leu Leu Ala Asn Lys Asp Phe 260 265

270 Val Lys Leu Arg Lys Leu Leu Ser Cys Asp Glu Thr Ala Leu Lys Gly 275 280 285 Ala Gln Gln Leu Ile Arg Gln Gln Asn Pro Lys Pro Gly Ser Glu Phe 290 295 300 Ala Thr Phe Ser His Asp His Phe Ile Gln His Asp Val Val Val Lys 305 310 315 320 Lys Ile Lys Gly Ile Trp Val Ala Ser Leu Asn Asp Gly Val Ile Pro 325 330 335 Lys Leu Arg Ile Asn Gln Leu Tyr Ala Asp Ile Leu Lys Arg Asn Arg 340 345 350 Glu Ser Ser Gly Gln Tyr Leu Gln Ser Gln Met Gln Glu Ala Lys Trp 355 360 365 Met Ile Lys Asn Ile Gln Gln Arg Phe Ser Thr Ile Leu Arg Val Ser 370 375 380 Gln Ala Ile Val Asp Arg Gln Arg Asn Phe Phe Glu His Gly Asp Ile 385 390 395 400 Ala Met Arg Pro Leu Val Leu Arg Glu Ile Ala Glu Glu Leu Asp Leu 405 410 415 His Glu Ser Thr Val Ser Arg Val Thr Thr His Lys Tyr Met Leu Thr 420 425 430 Pro Arg Gly Val Tyr Glu Leu Lys Tyr Phe Phe Gly Ser Ser Val Ala 435 440 445 Thr Asp Ala Gly Gly Ser Cys Ser Ala Thr Ala Ile Arg Ala Leu Ile 450 455 460 Lys Gln Met Val Ala Glu Glu Asn Pro Lys Lys Pro Leu Ser Asp Asn 465 470 475 480 Gln Ile Thr Asp Thr Leu Ala Gln Gln Gly Ile Val Val Ala Arg Arg 485 490 495 Thr Ile Ala Lys Tyr Arg Glu Ser Leu Asn Ile Pro Pro Ala Asn Leu 500 505 510 Arg Lys Ser Leu 515 19 1920 DNA Methylophilus methylotrophus CDS (1)..(1920) 19 atg gca aag att atc ggt att gat ttg gga acg aca aac tca tgc gtc 48 Met Ala Lys Ile Ile Gly Ile Asp Leu Gly Thr Thr Asn Ser Cys Val 1 5 10 15 gcc gtg atg gaa ggt ggc aag cca cgt gtg att gag aac gct gaa ggc 96 Ala Val Met Glu Gly Gly Lys Pro Arg Val Ile Glu Asn Ala Glu Gly 20 25 30 gcg cgt acc acg cct tct att att gct tac cag gaa gac ggc gaa att 144 Ala Arg Thr Thr Pro Ser Ile Ile Ala Tyr Gln Glu Asp Gly Glu Ile 35 40 45 ctg gtg ggt gca cca gca aaa cgt cag gca gtg act aac ccc aaa aat 192 Leu Val Gly Ala Pro Ala Lys Arg Gln Ala Val Thr Asn Pro Lys Asn 50 55 60 acc ctg ttt gcg gtg aag cgt ttg att ggc cgt cgc ttt gac gaa aaa 240 Thr Leu Phe Ala Val Lys Arg Leu Ile Gly Arg Arg Phe Asp Glu Lys 65 70 75 80 gaa gtg caa aaa gac atc gac ctc atg cct tac acc att gcc aag gct 288 Glu Val Gln Lys Asp Ile Asp Leu Met Pro Tyr Thr Ile Ala Lys Ala 85 90 95 gac aat ggt gac gca tgg gta gaa gtg cgt ggc aaa aaa cag gcg cca 336 Asp Asn Gly Asp Ala Trp Val Glu Val Arg Gly Lys Lys Gln Ala Pro 100 105 110 cca caa att tct gcc gaa gtg ttg cgc aaa atg aag aaa act gct gaa 384 Pro Gln Ile Ser Ala Glu Val Leu Arg Lys Met Lys Lys Thr Ala Glu 115 120 125 gac tac ctg ggc gaa gaa gtg acc gag gcc gtg att acc gtg ccc gct 432 Asp Tyr Leu Gly Glu Glu Val Thr Glu Ala Val Ile Thr Val Pro Ala 130 135 140 tac ttt aac gat agc cag cgt cag gca act aaa gat gca ggc cgt atc 480 Tyr Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp Ala Gly Arg Ile 145 150 155 160 gct ggt ctg gaa gta aaa cgt atc atc aac gag cca acg gca gcg gca 528 Ala Gly Leu Glu Val Lys Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala 165 170 175 ctg gca ttt ggt ttg gac aaa cag gaa ggt gac cgc aag att gcg gta 576 Leu Ala Phe Gly Leu Asp Lys Gln Glu Gly Asp Arg Lys Ile Ala Val 180 185 190 tat gac ctg ggt ggc ggc act ttc gat att tcg att att gaa att act 624 Tyr Asp Leu Gly Gly Gly Thr Phe Asp Ile Ser Ile Ile Glu Ile Thr 195 200 205 gaa atc gac ggc gag cac cag ttc gaa gtg ttg tct acc aat ggt gac 672 Glu Ile Asp Gly Glu His Gln Phe Glu Val Leu Ser Thr Asn Gly Asp 210 215 220 aca ttc ctc ggt ggt gaa gac ttt gat aac cgc atc atc gac ttt ttg 720 Thr Phe Leu Gly Gly Glu Asp Phe Asp Asn Arg Ile Ile Asp Phe Leu 225 230 235 240 gct gac gaa ttc aag aaa gaa aac ggc ttg gac ttg cgt aac gac ttg 768 Ala Asp Glu Phe Lys Lys Glu Asn Gly Leu Asp Leu Arg Asn Asp Leu 245 250 255 ctg gca aaa cag cgt ctg aaa gag gcg gcc gaa aaa gcc aag atc gaa 816 Leu Ala Lys Gln Arg Leu Lys Glu Ala Ala Glu Lys Ala Lys Ile Glu 260 265 270 ttg tct ggt gca cag cag act gaa gtg aac ctg ccg tac atc acg gct 864 Leu Ser Gly Ala Gln Gln Thr Glu Val Asn Leu Pro Tyr Ile Thr Ala 275 280 285 gat gcg acc ggt cct aag cac ttg gtg gtg aaa atc act cgt gcc aag 912 Asp Ala Thr Gly Pro Lys His Leu Val Val Lys Ile Thr Arg Ala Lys 290 295 300 ctg gag tca ttg gtg gaa gac ctg att gag cga acc atc agc cca tgt 960 Leu Glu Ser Leu Val Glu Asp Leu Ile Glu Arg Thr Ile Ser Pro Cys 305 310 315 320 aaa acg gca ttg aaa gat gca ggc gtg tct cct tca gat att tct gac 1008 Lys Thr Ala Leu Lys Asp Ala Gly Val Ser Pro Ser Asp Ile Ser Asp 325 330 335 gtg att ttg gtg ggc ggt cag agc cgg atg cct aaa gtg caa gaa aaa 1056 Val Ile Leu Val Gly Gly Gln Ser Arg Met Pro Lys Val Gln Glu Lys 340 345 350 gtg aaa gag att ttt ggc aag gaa cca cgt aaa gat gtg aac ccg gat 1104 Val Lys Glu Ile Phe Gly Lys Glu Pro Arg Lys Asp Val Asn Pro Asp 355 360 365 gaa gcc gtg gcc gta ggt gct gcg atc cag ggc ggc gta ctg aaa ggc 1152 Glu Ala Val Ala Val Gly Ala Ala Ile Gln Gly Gly Val Leu Lys Gly 370 375 380 gac gtg aaa gac gtg ttg ctg ctg gac gtg aca cca ttg tcc ctg ggt 1200 Asp Val Lys Asp Val Leu Leu Leu Asp Val Thr Pro Leu Ser Leu Gly 385 390 395 400 att gaa acc ctg ggt agc gtg atg acc aag ctg atc aag aaa aac acc 1248 Ile Glu Thr Leu Gly Ser Val Met Thr Lys Leu Ile Lys Lys Asn Thr 405 410 415 acg att cct acc aag gca tca caa gtg ttc tcg aca gct gaa gac aac 1296 Thr Ile Pro Thr Lys Ala Ser Gln Val Phe Ser Thr Ala Glu Asp Asn 420 425 430 cag aat gcg gtg act atc cat gtg ttg caa ggt gaa cgc gaa atg gct 1344 Gln Asn Ala Val Thr Ile His Val Leu Gln Gly Glu Arg Glu Met Ala 435 440 445 tcc ggc aac aag agc ctg ggc cag ttt aac ctc agc gac att cca cct 1392 Ser Gly Asn Lys Ser Leu Gly Gln Phe Asn Leu Ser Asp Ile Pro Pro 450 455 460 gca ccg cgt ggc atg ccg caa att gaa gtg act ttt gat atc gat gcc 1440 Ala Pro Arg Gly Met Pro Gln Ile Glu Val Thr Phe Asp Ile Asp Ala 465 470 475 480 aac ggt att ttg cat gtg tct gcc aaa gac aag gcc act ggc aaa gaa 1488 Asn Gly Ile Leu His Val Ser Ala Lys Asp Lys Ala Thr Gly Lys Glu 485 490 495 aac aag atc acc atc aaa gct aac tcc ggc ttg tct gaa gaa gaa att 1536 Asn Lys Ile Thr Ile Lys Ala Asn Ser Gly Leu Ser Glu Glu Glu Ile 500 505 510 cag cgc atg gaa gaa gat gcg gct aaa tat gct gac gaa gac aaa aaa 1584 Gln Arg Met Glu Glu Asp Ala Ala Lys Tyr Ala Asp Glu Asp Lys Lys 515 520 525 ctg cgt gaa ctg gtc gac gcc cgt aac cag gcg gac agc gtg ttg cac 1632 Leu Arg Glu Leu Val Asp Ala Arg Asn Gln Ala Asp Ser Val Leu His 530 535 540 agc gtg aaa aaa tcg ttg gct gag cat ggc gac aag att gag gcc gat 1680 Ser Val Lys Lys Ser Leu Ala Glu His Gly Asp Lys Ile Glu Ala Asp 545 550 555 560 gaa aaa gcc aag att gaa gac gca att aaa gac ctg gaa gcg gta gct 1728 Glu Lys Ala Lys Ile Glu Asp Ala Ile Lys Asp Leu Glu Ala Val Ala 565 570 575 aaa gac ggc gac gat aaa gaa gtg att gaa gcc aag acc aat gct ttg 1776 Lys Asp Gly Asp Asp Lys Glu Val Ile Glu Ala Lys Thr Asn Ala Leu 580 585 590 atg gaa gct tca caa aaa ctg ggt gaa aag gtc tat gcc gag caa cag 1824 Met Glu Ala Ser Gln Lys Leu Gly Glu Lys Val Tyr Ala Glu Gln Gln 595 600 605 gca cag gcc aat acc gag agt gca caa gct gaa act gaa aaa aca gtg 1872 Ala Gln Ala Asn Thr Glu Ser Ala Gln Ala Glu Thr Glu Lys Thr Val 610 615 620 gaa ggc gat gtg gtg gat gcc gag ttt gaa gaa gtg aag aaa aac taa 1920 Glu Gly Asp Val Val Asp Ala Glu Phe Glu Glu Val Lys Lys Asn 625 630 635 20 639 PRT Methylophilus methylotrophus 20 Met Ala Lys Ile Ile Gly Ile Asp Leu Gly Thr Thr Asn Ser Cys Val 1 5 10 15 Ala Val Met Glu Gly Gly Lys Pro Arg Val Ile Glu Asn Ala Glu Gly 20 25 30 Ala Arg Thr Thr Pro Ser Ile Ile Ala Tyr Gln Glu Asp Gly Glu Ile 35 40 45 Leu Val Gly Ala Pro Ala Lys Arg Gln Ala Val Thr Asn Pro Lys Asn 50 55 60 Thr Leu Phe Ala Val Lys Arg Leu Ile Gly Arg Arg Phe Asp Glu Lys 65 70 75 80 Glu Val Gln Lys Asp Ile Asp Leu Met Pro Tyr Thr Ile Ala Lys Ala 85 90 95 Asp Asn Gly Asp Ala Trp Val Glu Val Arg Gly Lys Lys Gln Ala Pro 100 105 110 Pro Gln Ile Ser Ala Glu Val Leu Arg Lys Met Lys Lys Thr Ala Glu 115 120 125 Asp Tyr Leu Gly Glu Glu Val Thr Glu Ala Val Ile Thr Val Pro Ala 130 135 140 Tyr Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp Ala Gly Arg Ile 145 150 155 160 Ala Gly Leu Glu Val Lys Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala 165 170 175 Leu Ala Phe Gly Leu Asp Lys Gln Glu Gly Asp Arg Lys Ile Ala Val 180 185 190 Tyr Asp Leu Gly Gly Gly Thr Phe Asp Ile Ser Ile Ile Glu Ile Thr 195 200 205 Glu Ile Asp Gly Glu His Gln Phe Glu Val Leu Ser Thr Asn Gly Asp 210 215 220 Thr Phe Leu Gly Gly Glu Asp Phe Asp Asn Arg Ile Ile Asp Phe Leu 225 230 235 240 Ala Asp Glu Phe Lys Lys Glu Asn Gly Leu Asp Leu Arg Asn Asp Leu 245 250 255 Leu Ala Lys Gln Arg Leu Lys Glu Ala Ala Glu Lys Ala Lys Ile Glu 260 265 270 Leu Ser Gly Ala Gln Gln Thr Glu Val Asn Leu Pro Tyr Ile Thr Ala 275 280 285 Asp Ala Thr Gly Pro Lys His Leu Val Val Lys Ile Thr Arg Ala Lys 290 295 300 Leu Glu Ser Leu Val Glu Asp Leu Ile Glu Arg Thr Ile Ser Pro Cys 305 310 315 320 Lys Thr Ala Leu Lys Asp Ala Gly Val Ser Pro Ser Asp Ile Ser Asp 325 330 335 Val Ile Leu Val Gly Gly Gln Ser Arg Met Pro Lys Val Gln Glu Lys 340 345 350 Val Lys Glu Ile Phe Gly Lys Glu Pro Arg Lys Asp Val Asn Pro Asp 355 360 365 Glu Ala Val Ala Val Gly Ala Ala Ile Gln Gly Gly Val Leu Lys Gly 370 375 380 Asp Val Lys Asp Val Leu Leu Leu Asp Val Thr Pro Leu Ser Leu Gly 385 390 395 400 Ile Glu Thr Leu Gly Ser Val Met Thr Lys Leu Ile Lys Lys Asn Thr 405 410 415 Thr Ile Pro Thr Lys Ala Ser Gln Val Phe Ser Thr Ala Glu Asp Asn 420 425 430 Gln Asn Ala Val Thr Ile His Val Leu Gln Gly Glu Arg Glu Met Ala 435 440 445 Ser Gly Asn Lys Ser Leu Gly Gln Phe Asn Leu Ser Asp Ile Pro Pro 450 455 460 Ala Pro Arg Gly Met Pro Gln Ile Glu Val Thr Phe Asp Ile Asp Ala 465 470 475 480 Asn Gly Ile Leu His Val Ser Ala Lys Asp Lys Ala Thr Gly Lys Glu 485 490 495 Asn Lys Ile Thr Ile Lys Ala Asn Ser Gly Leu Ser Glu Glu Glu Ile 500 505 510 Gln Arg Met Glu Glu Asp Ala Ala Lys Tyr Ala Asp Glu Asp Lys Lys 515 520 525 Leu Arg Glu Leu Val Asp Ala Arg Asn Gln Ala Asp Ser Val Leu His 530 535 540 Ser Val Lys Lys Ser Leu Ala Glu His Gly Asp Lys Ile Glu Ala Asp 545 550 555 560 Glu Lys Ala Lys Ile Glu Asp Ala Ile Lys Asp Leu Glu Ala Val Ala 565 570 575 Lys Asp Gly Asp Asp Lys Glu Val Ile Glu Ala Lys Thr Asn Ala Leu 580 585 590 Met Glu Ala Ser Gln Lys Leu Gly Glu Lys Val Tyr Ala Glu Gln Gln 595 600 605 Ala Gln Ala Asn Thr Glu Ser Ala Gln Ala Glu Thr Glu Lys Thr Val 610 615 620 Glu Gly Asp Val Val Asp Ala Glu Phe Glu Glu Val Lys Lys Asn 625 630 635 21 1131 DNA Methylophilus methylotrophus CDS (1)..(1131) 21 atg gca gct gca aaa aaa gat tat tac gaa gtg tta ggt gta aac cgc 48 Met Ala Ala Ala Lys Lys Asp Tyr Tyr Glu Val Leu Gly Val Asn Arg 1 5 10 15 gac gcc agc gaa gag gaa att aaa aaa gcc ttt aaa aaa ctg gcc atg 96 Asp Ala Ser Glu Glu Glu Ile Lys Lys Ala Phe Lys Lys Leu Ala Met 20 25 30 aag ttt cac ccg gac cgc aac ccg gat aac ccc aaa gcc gaa gaa agc 144 Lys Phe His Pro Asp Arg Asn Pro Asp Asn Pro Lys Ala Glu Glu Ser 35 40 45 ttt aag gaa gcc aaa gag gct tat gag ata ttg agt gac gag cag aag 192 Phe Lys Glu Ala Lys Glu Ala Tyr Glu Ile Leu Ser Asp Glu Gln Lys 50 55 60 cgc gct gct tat gac caa tat ggc cat gcc ggc gta gac ccc agc atg 240 Arg Ala Ala Tyr Asp Gln Tyr Gly His Ala Gly Val Asp Pro Ser Met 65 70 75 80 ggc ggt ggc ggt gga ttt ggc gga ttc aat tct ggc aac ttc agt gac 288 Gly Gly Gly Gly Gly Phe Gly Gly Phe Asn Ser Gly Asn Phe Ser Asp 85 90 95 gcg ttt ggc gat att ttt ggg gat att ttc ggt ggt gcg cgc aac cag 336 Ala Phe Gly Asp Ile Phe Gly Asp Ile Phe Gly Gly Ala Arg Asn Gln 100 105 110 cgc tcg aat gtg tac cgt ggt gcg gat tta cgt tac aac ctg gaa att 384 Arg Ser Asn Val Tyr Arg Gly Ala Asp Leu Arg Tyr Asn Leu Glu Ile 115 120 125 tcg ctg gaa gat gcg gcc aaa ggg act gag acc aaa atc cgt att ccg 432 Ser Leu Glu Asp Ala Ala Lys Gly Thr Glu Thr Lys Ile Arg Ile Pro 130 135 140 gta cag acc act tgt gaa acc tgt cac ggt tca ggc gcg cgt cca ggg 480 Val Gln Thr Thr Cys Glu Thr Cys His Gly Ser Gly Ala Arg Pro Gly 145 150 155 160 aca tcc cct gtg act tgt acc acc tgt aac ggt cac ggc cag gta cgt 528 Thr Ser Pro Val Thr Cys Thr Thr Cys Asn Gly His Gly Gln Val Arg 165 170 175 atg caa cag ggt ttt ttc tct gta cag caa acc tgt ccc aaa tgt cat 576 Met Gln Gln Gly Phe Phe Ser Val Gln Gln Thr Cys Pro Lys Cys His 180 185 190 ggc acc ggc aaa atg gtg aaa gag cct tgc cca act tgc cat ggc ggt 624 Gly Thr Gly Lys Met Val Lys Glu Pro Cys Pro Thr Cys His Gly Gly 195 200 205 ggt cgc gtc aaa cag aat aaa acg ctg aat gtg aag att cca gcg ggt 672 Gly Arg Val Lys Gln Asn Lys Thr Leu Asn Val Lys Ile Pro Ala Gly 210 215 220 gtc gac gag ggg gat cgt atc cgc ctc agc ggt gag ggt gaa gct ggc 720 Val Asp Glu Gly Asp Arg Ile Arg Leu Ser Gly Glu Gly Glu Ala Gly 225 230 235 240 gtc aat ggc ggc cca acg ggt gat ttg tat gtg gtg gtg cat ctc aag 768 Val Asn Gly Gly Pro Thr Gly Asp Leu Tyr Val Val Val His Leu Lys 245 250 255 gaa cac ccg att ttc cag cgc gaa ggt gca aac ctg cat tgt gaa atg 816 Glu His Pro Ile Phe Gln Arg Glu Gly Ala Asn Leu His Cys Glu Met 260 265 270 cct atc agc ttt agt acc gcg gcc tta ggc ggc gaa att gaa gtg ccg 864 Pro Ile Ser Phe Ser Thr Ala Ala Leu Gly Gly Glu Ile Glu Val Pro 275 280 285 acg ctg gat ggt gca gcc aag atg aag ata ccg gct gaa acg caa aca 912 Thr Leu Asp Gly Ala Ala Lys Met Lys Ile Pro Ala Glu Thr Gln Thr 290 295 300 ggc agt gta ttc cgt ttg cgc ggc aag ggt atc aag ccc tta cgc tcc 960 Gly Ser Val Phe Arg Leu Arg Gly Lys Gly Ile Lys Pro Leu Arg Ser 305 310 315 320 agc gaa tat ggt gat ttg atg gtg cat gtg

gtc gtt gaa acg cca gtg 1008 Ser Glu Tyr Gly Asp Leu Met Val His Val Val Val Glu Thr Pro Val 325 330 335 cgc ctg aca gaa aag cag aaa gaa ctg ttg cgt gag ttt gaa agt agt 1056 Arg Leu Thr Glu Lys Gln Lys Glu Leu Leu Arg Glu Phe Glu Ser Ser 340 345 350 act cag gca gat gcg gga aaa cat agc ccc aag aat aaa agc tgg gta 1104 Thr Gln Ala Asp Ala Gly Lys His Ser Pro Lys Asn Lys Ser Trp Val 355 360 365 gat aaa gcc cgc gat ttt ttt agc tag 1131 Asp Lys Ala Arg Asp Phe Phe Ser 370 375 22 376 PRT Methylophilus methylotrophus 22 Met Ala Ala Ala Lys Lys Asp Tyr Tyr Glu Val Leu Gly Val Asn Arg 1 5 10 15 Asp Ala Ser Glu Glu Glu Ile Lys Lys Ala Phe Lys Lys Leu Ala Met 20 25 30 Lys Phe His Pro Asp Arg Asn Pro Asp Asn Pro Lys Ala Glu Glu Ser 35 40 45 Phe Lys Glu Ala Lys Glu Ala Tyr Glu Ile Leu Ser Asp Glu Gln Lys 50 55 60 Arg Ala Ala Tyr Asp Gln Tyr Gly His Ala Gly Val Asp Pro Ser Met 65 70 75 80 Gly Gly Gly Gly Gly Phe Gly Gly Phe Asn Ser Gly Asn Phe Ser Asp 85 90 95 Ala Phe Gly Asp Ile Phe Gly Asp Ile Phe Gly Gly Ala Arg Asn Gln 100 105 110 Arg Ser Asn Val Tyr Arg Gly Ala Asp Leu Arg Tyr Asn Leu Glu Ile 115 120 125 Ser Leu Glu Asp Ala Ala Lys Gly Thr Glu Thr Lys Ile Arg Ile Pro 130 135 140 Val Gln Thr Thr Cys Glu Thr Cys His Gly Ser Gly Ala Arg Pro Gly 145 150 155 160 Thr Ser Pro Val Thr Cys Thr Thr Cys Asn Gly His Gly Gln Val Arg 165 170 175 Met Gln Gln Gly Phe Phe Ser Val Gln Gln Thr Cys Pro Lys Cys His 180 185 190 Gly Thr Gly Lys Met Val Lys Glu Pro Cys Pro Thr Cys His Gly Gly 195 200 205 Gly Arg Val Lys Gln Asn Lys Thr Leu Asn Val Lys Ile Pro Ala Gly 210 215 220 Val Asp Glu Gly Asp Arg Ile Arg Leu Ser Gly Glu Gly Glu Ala Gly 225 230 235 240 Val Asn Gly Gly Pro Thr Gly Asp Leu Tyr Val Val Val His Leu Lys 245 250 255 Glu His Pro Ile Phe Gln Arg Glu Gly Ala Asn Leu His Cys Glu Met 260 265 270 Pro Ile Ser Phe Ser Thr Ala Ala Leu Gly Gly Glu Ile Glu Val Pro 275 280 285 Thr Leu Asp Gly Ala Ala Lys Met Lys Ile Pro Ala Glu Thr Gln Thr 290 295 300 Gly Ser Val Phe Arg Leu Arg Gly Lys Gly Ile Lys Pro Leu Arg Ser 305 310 315 320 Ser Glu Tyr Gly Asp Leu Met Val His Val Val Val Glu Thr Pro Val 325 330 335 Arg Leu Thr Glu Lys Gln Lys Glu Leu Leu Arg Glu Phe Glu Ser Ser 340 345 350 Thr Gln Ala Asp Ala Gly Lys His Ser Pro Lys Asn Lys Ser Trp Val 355 360 365 Asp Lys Ala Arg Asp Phe Phe Ser 370 375 23 1890 DNA Methylophilus methylotrophus CDS (1)..(1890) 23 atg gcc tta tta cag att tct gaa ccc gga caa tcc cca gca ccc cat 48 Met Ala Leu Leu Gln Ile Ser Glu Pro Gly Gln Ser Pro Ala Pro His 1 5 10 15 cag cat aag ctg gcc att ggc ata gac ctg ggt acg acc aac tcc ctg 96 Gln His Lys Leu Ala Ile Gly Ile Asp Leu Gly Thr Thr Asn Ser Leu 20 25 30 gtc gcc act gtg cgt agc ggc atg agt aca gtg ttg cat gac gaa cat 144 Val Ala Thr Val Arg Ser Gly Met Ser Thr Val Leu His Asp Glu His 35 40 45 ggt cat gca ttg cta cct tct gtt gtg cgt tac ctc aat gac gcc gta 192 Gly His Ala Leu Leu Pro Ser Val Val Arg Tyr Leu Asn Asp Ala Val 50 55 60 att gtc ggt cac gag gcg cag gct gca caa agc cag gac ccg gtg aat 240 Ile Val Gly His Glu Ala Gln Ala Ala Gln Ser Gln Asp Pro Val Asn 65 70 75 80 acc atc gtg tcg gtc aaa cgc ttt atg ggg cgt gca ttg cat gac atc 288 Thr Ile Val Ser Val Lys Arg Phe Met Gly Arg Ala Leu His Asp Ile 85 90 95 acc gat aga gcg cat atc ccc tac cac ttt gtt gaa aat gac agc tca 336 Thr Asp Arg Ala His Ile Pro Tyr His Phe Val Glu Asn Asp Ser Ser 100 105 110 caa ggc atg ctg gaa ctc aaa acc cgc gca ggg ttg aaa agc ccg gtg 384 Gln Gly Met Leu Glu Leu Lys Thr Arg Ala Gly Leu Lys Ser Pro Val 115 120 125 gag att tct gcc gaa atc ctc aaa aca ctc aaa gcc cgt gcc gaa aaa 432 Glu Ile Ser Ala Glu Ile Leu Lys Thr Leu Lys Ala Arg Ala Glu Lys 130 135 140 gct tta ggc ggt gaa ctc acg ggg gcc gtc att acc gtc ccc gcc tat 480 Ala Leu Gly Gly Glu Leu Thr Gly Ala Val Ile Thr Val Pro Ala Tyr 145 150 155 160 ttt gac gat gcc cag cgc cag gcc acc aaa gat gct gca cgc ctc gca 528 Phe Asp Asp Ala Gln Arg Gln Ala Thr Lys Asp Ala Ala Arg Leu Ala 165 170 175 ggg ttg cat gta ttg cgc ctg ctc aat gag ccg acg gca gcg gca gtc 576 Gly Leu His Val Leu Arg Leu Leu Asn Glu Pro Thr Ala Ala Ala Val 180 185 190 gcc tat ggt ctg gat aat gcc gcc gaa ggt gtc tat gtc att tat gac 624 Ala Tyr Gly Leu Asp Asn Ala Ala Glu Gly Val Tyr Val Ile Tyr Asp 195 200 205 ctg ggt ggt ggc acc ttt gat att tct atc tta cgc ttg agc aaa ggc 672 Leu Gly Gly Gly Thr Phe Asp Ile Ser Ile Leu Arg Leu Ser Lys Gly 210 215 220 gta ttt gag gta ctg gcc acc aat ggc gac tca gcc ctg ggg gga gat 720 Val Phe Glu Val Leu Ala Thr Asn Gly Asp Ser Ala Leu Gly Gly Asp 225 230 235 240 gac ttc gat cat cgc att tat tgc tgg gta ctg gac cag gta cgc agc 768 Asp Phe Asp His Arg Ile Tyr Cys Trp Val Leu Asp Gln Val Arg Ser 245 250 255 aag acc aaa gac ttc aaa ccg ctc acc gaa gaa gat aca cgc ctg ttg 816 Lys Thr Lys Asp Phe Lys Pro Leu Thr Glu Glu Asp Thr Arg Leu Leu 260 265 270 ctg act aag tca cgc cag gcc aaa gaa tgg ctg aca gat aac cac gaa 864 Leu Thr Lys Ser Arg Gln Ala Lys Glu Trp Leu Thr Asp Asn His Glu 275 280 285 gcc aat att gtc tgc aaa ctg agt aat ggt gca ctg gtt gat gaa acc 912 Ala Asn Ile Val Cys Lys Leu Ser Asn Gly Ala Leu Val Asp Glu Thr 290 295 300 ttg act gac agc cag ttt gtc gca ctg act gaa cac ctg gtc atc aag 960 Leu Thr Asp Ser Gln Phe Val Ala Leu Thr Glu His Leu Val Ile Lys 305 310 315 320 acg ctc agc cct acc cgc aaa gcc atg cgt gat gca ggc ctg aac att 1008 Thr Leu Ser Pro Thr Arg Lys Ala Met Arg Asp Ala Gly Leu Asn Ile 325 330 335 gca gaa atc aaa ggc gtg gta ctg gtc ggt ggc gcc acc cgt atg ccg 1056 Ala Glu Ile Lys Gly Val Val Leu Val Gly Gly Ala Thr Arg Met Pro 340 345 350 cat atc cgc cat gcg gta cag gct ttt ttc gag cag gaa ccg ctg acc 1104 His Ile Arg His Ala Val Gln Ala Phe Phe Glu Gln Glu Pro Leu Thr 355 360 365 aac ctt gac ccg gat aaa gtc gtg gca ctc ggt gcc gct atc cag gcc 1152 Asn Leu Asp Pro Asp Lys Val Val Ala Leu Gly Ala Ala Ile Gln Ala 370 375 380 aat gtg ctg gca gga aac cgc agt gac gaa gaa tta ctg tta ctg gac 1200 Asn Val Leu Ala Gly Asn Arg Ser Asp Glu Glu Leu Leu Leu Leu Asp 385 390 395 400 gtg acc ccg ctc tca ctg ggg ctg gaa acc atg ggc gga ctg gtt gaa 1248 Val Thr Pro Leu Ser Leu Gly Leu Glu Thr Met Gly Gly Leu Val Glu 405 410 415 aaa gtt att ccg cgc aac tct aca ctg ccg att gca cgt gca cag gac 1296 Lys Val Ile Pro Arg Asn Ser Thr Leu Pro Ile Ala Arg Ala Gln Asp 420 425 430 ttc acc act tac aag gat ggc cag acc gcc atg gcc att cat gtt gtg 1344 Phe Thr Thr Tyr Lys Asp Gly Gln Thr Ala Met Ala Ile His Val Val 435 440 445 caa ggc gag cgt gaa ctg gtc agt gac tgc cgc tcg ctg gca cgc ttt 1392 Gln Gly Glu Arg Glu Leu Val Ser Asp Cys Arg Ser Leu Ala Arg Phe 450 455 460 gaa ctg cgt ggc att ccc cca atg gca gcc ggc gca gcg cgt atc cgc 1440 Glu Leu Arg Gly Ile Pro Pro Met Ala Ala Gly Ala Ala Arg Ile Arg 465 470 475 480 gtc act ttc cag gta gat gct gat ggg ctg cta tca gtc agt gca cgc 1488 Val Thr Phe Gln Val Asp Ala Asp Gly Leu Leu Ser Val Ser Ala Arg 485 490 495 gaa caa acc agc ggc ata gag gcc aac atc acg gtc aag ccc tct tat 1536 Glu Gln Thr Ser Gly Ile Glu Ala Asn Ile Thr Val Lys Pro Ser Tyr 500 505 510 ggc ctc agt gaa gac cag att agc ggt atg ctg aaa gat tcg ttt ggt 1584 Gly Leu Ser Glu Asp Gln Ile Ser Gly Met Leu Lys Asp Ser Phe Gly 515 520 525 gct gct gag agt gac aaa caa gca cgc atg ctg cgt gaa gct gtt gtg 1632 Ala Ala Glu Ser Asp Lys Gln Ala Arg Met Leu Arg Glu Ala Val Val 530 535 540 gat gct caa cgc ctg gtc gag gcg att caa gca gca ctg gca gaa gat 1680 Asp Ala Gln Arg Leu Val Glu Ala Ile Gln Ala Ala Leu Ala Glu Asp 545 550 555 560 gga gag acg tta cta tct gct gac gaa cgt cag cag ata aac gcc cat 1728 Gly Glu Thr Leu Leu Ser Ala Asp Glu Arg Gln Gln Ile Asn Ala His 565 570 575 ata gac acc tta ttg gcc ttg tgc cag ggc gac gat agc cag gcc gtt 1776 Ile Asp Thr Leu Leu Ala Leu Cys Gln Gly Asp Asp Ser Gln Ala Val 580 585 590 aaa cag gct act gaa gca ctg aac cat gcc acc gag gca ttt gct gcc 1824 Lys Gln Ala Thr Glu Ala Leu Asn His Ala Thr Glu Ala Phe Ala Ala 595 600 605 aag cgc atg gat gcc tcg gta caa aaa gcc ctg gcc ggt aaa aac ctg 1872 Lys Arg Met Asp Ala Ser Val Gln Lys Ala Leu Ala Gly Lys Asn Leu 610 615 620 gat tca ctg gaa cta tga 1890 Asp Ser Leu Glu Leu 625 24 629 PRT Methylophilus methylotrophus 24 Met Ala Leu Leu Gln Ile Ser Glu Pro Gly Gln Ser Pro Ala Pro His 1 5 10 15 Gln His Lys Leu Ala Ile Gly Ile Asp Leu Gly Thr Thr Asn Ser Leu 20 25 30 Val Ala Thr Val Arg Ser Gly Met Ser Thr Val Leu His Asp Glu His 35 40 45 Gly His Ala Leu Leu Pro Ser Val Val Arg Tyr Leu Asn Asp Ala Val 50 55 60 Ile Val Gly His Glu Ala Gln Ala Ala Gln Ser Gln Asp Pro Val Asn 65 70 75 80 Thr Ile Val Ser Val Lys Arg Phe Met Gly Arg Ala Leu His Asp Ile 85 90 95 Thr Asp Arg Ala His Ile Pro Tyr His Phe Val Glu Asn Asp Ser Ser 100 105 110 Gln Gly Met Leu Glu Leu Lys Thr Arg Ala Gly Leu Lys Ser Pro Val 115 120 125 Glu Ile Ser Ala Glu Ile Leu Lys Thr Leu Lys Ala Arg Ala Glu Lys 130 135 140 Ala Leu Gly Gly Glu Leu Thr Gly Ala Val Ile Thr Val Pro Ala Tyr 145 150 155 160 Phe Asp Asp Ala Gln Arg Gln Ala Thr Lys Asp Ala Ala Arg Leu Ala 165 170 175 Gly Leu His Val Leu Arg Leu Leu Asn Glu Pro Thr Ala Ala Ala Val 180 185 190 Ala Tyr Gly Leu Asp Asn Ala Ala Glu Gly Val Tyr Val Ile Tyr Asp 195 200 205 Leu Gly Gly Gly Thr Phe Asp Ile Ser Ile Leu Arg Leu Ser Lys Gly 210 215 220 Val Phe Glu Val Leu Ala Thr Asn Gly Asp Ser Ala Leu Gly Gly Asp 225 230 235 240 Asp Phe Asp His Arg Ile Tyr Cys Trp Val Leu Asp Gln Val Arg Ser 245 250 255 Lys Thr Lys Asp Phe Lys Pro Leu Thr Glu Glu Asp Thr Arg Leu Leu 260 265 270 Leu Thr Lys Ser Arg Gln Ala Lys Glu Trp Leu Thr Asp Asn His Glu 275 280 285 Ala Asn Ile Val Cys Lys Leu Ser Asn Gly Ala Leu Val Asp Glu Thr 290 295 300 Leu Thr Asp Ser Gln Phe Val Ala Leu Thr Glu His Leu Val Ile Lys 305 310 315 320 Thr Leu Ser Pro Thr Arg Lys Ala Met Arg Asp Ala Gly Leu Asn Ile 325 330 335 Ala Glu Ile Lys Gly Val Val Leu Val Gly Gly Ala Thr Arg Met Pro 340 345 350 His Ile Arg His Ala Val Gln Ala Phe Phe Glu Gln Glu Pro Leu Thr 355 360 365 Asn Leu Asp Pro Asp Lys Val Val Ala Leu Gly Ala Ala Ile Gln Ala 370 375 380 Asn Val Leu Ala Gly Asn Arg Ser Asp Glu Glu Leu Leu Leu Leu Asp 385 390 395 400 Val Thr Pro Leu Ser Leu Gly Leu Glu Thr Met Gly Gly Leu Val Glu 405 410 415 Lys Val Ile Pro Arg Asn Ser Thr Leu Pro Ile Ala Arg Ala Gln Asp 420 425 430 Phe Thr Thr Tyr Lys Asp Gly Gln Thr Ala Met Ala Ile His Val Val 435 440 445 Gln Gly Glu Arg Glu Leu Val Ser Asp Cys Arg Ser Leu Ala Arg Phe 450 455 460 Glu Leu Arg Gly Ile Pro Pro Met Ala Ala Gly Ala Ala Arg Ile Arg 465 470 475 480 Val Thr Phe Gln Val Asp Ala Asp Gly Leu Leu Ser Val Ser Ala Arg 485 490 495 Glu Gln Thr Ser Gly Ile Glu Ala Asn Ile Thr Val Lys Pro Ser Tyr 500 505 510 Gly Leu Ser Glu Asp Gln Ile Ser Gly Met Leu Lys Asp Ser Phe Gly 515 520 525 Ala Ala Glu Ser Asp Lys Gln Ala Arg Met Leu Arg Glu Ala Val Val 530 535 540 Asp Ala Gln Arg Leu Val Glu Ala Ile Gln Ala Ala Leu Ala Glu Asp 545 550 555 560 Gly Glu Thr Leu Leu Ser Ala Asp Glu Arg Gln Gln Ile Asn Ala His 565 570 575 Ile Asp Thr Leu Leu Ala Leu Cys Gln Gly Asp Asp Ser Gln Ala Val 580 585 590 Lys Gln Ala Thr Glu Ala Leu Asn His Ala Thr Glu Ala Phe Ala Ala 595 600 605 Lys Arg Met Asp Ala Ser Val Gln Lys Ala Leu Ala Gly Lys Asn Leu 610 615 620 Asp Ser Leu Glu Leu 625 25 513 DNA Methylophilus methylotrophus CDS (1)..(513) 25 atg caa aac tac ttc gca ctg ttt caa ctg cca cag caa ttt gag tta 48 Met Gln Asn Tyr Phe Ala Leu Phe Gln Leu Pro Gln Gln Phe Glu Leu 1 5 10 15 gat cta act caa ctg gat agc cag tac cgt aaa tta caa gcc gaa gtg 96 Asp Leu Thr Gln Leu Asp Ser Gln Tyr Arg Lys Leu Gln Ala Glu Val 20 25 30 cat ccc gat aaa ttt gtg agt gcc tca cct gcc gaa cgc atg cac tca 144 His Pro Asp Lys Phe Val Ser Ala Ser Pro Ala Glu Arg Met His Ser 35 40 45 atg cag atg gcg acg ctg gca aat gaa gct tac caa acg ctt aaa cac 192 Met Gln Met Ala Thr Leu Ala Asn Glu Ala Tyr Gln Thr Leu Lys His 50 55 60 ccc acc gcc cgt gca cgc tac ctg ttg caa ctc cag ggt atc act aca 240 Pro Thr Ala Arg Ala Arg Tyr Leu Leu Gln Leu Gln Gly Ile Thr Thr 65 70 75 80 gat gaa gaa aac aac act gcc atg ccc gct gat ttt tta atg gca caa 288 Asp Glu Glu Asn Asn Thr Ala Met Pro Ala Asp Phe Leu Met Ala Gln 85 90 95 atg gag tgg cgc gag gcc att gac gat gcc aag tac agc aag gat atc 336 Met Glu Trp Arg Glu Ala Ile Asp Asp Ala Lys Tyr Ser Lys Asp Ile 100 105 110 gct gca ctg gat acc tta ttg aag gat atg cgc gcg caa gcc aca act 384 Ala Ala Leu Asp Thr Leu Leu Lys Asp Met Arg Ala Gln Ala Thr Thr 115 120 125 ttg cag cag caa gtc gca aca gag atc gag aca gcg ccc acc cta gct 432 Leu Gln Gln Gln Val Ala Thr Glu Ile Glu Thr Ala Pro Thr Leu Ala 130 135 140 gca ctc acc gta cgc aaa ttg cgt ttt att gat aaa gtg agt gaa gat 480 Ala Leu Thr Val Arg Lys Leu Arg Phe Ile Asp Lys Val Ser Glu Asp 145 150 155 160 gtg aat caa ctg atc gct cag ttg gaa gat tag 513 Val Asn Gln Leu Ile Ala Gln Leu Glu Asp 165 170 26 170 PRT Methylophilus methylotrophus 26 Met Gln Asn Tyr Phe Ala Leu Phe Gln Leu Pro Gln Gln Phe Glu Leu 1 5 10 15 Asp Leu Thr Gln Leu Asp Ser Gln Tyr Arg Lys Leu Gln Ala Glu Val 20 25

30 His Pro Asp Lys Phe Val Ser Ala Ser Pro Ala Glu Arg Met His Ser 35 40 45 Met Gln Met Ala Thr Leu Ala Asn Glu Ala Tyr Gln Thr Leu Lys His 50 55 60 Pro Thr Ala Arg Ala Arg Tyr Leu Leu Gln Leu Gln Gly Ile Thr Thr 65 70 75 80 Asp Glu Glu Asn Asn Thr Ala Met Pro Ala Asp Phe Leu Met Ala Gln 85 90 95 Met Glu Trp Arg Glu Ala Ile Asp Asp Ala Lys Tyr Ser Lys Asp Ile 100 105 110 Ala Ala Leu Asp Thr Leu Leu Lys Asp Met Arg Ala Gln Ala Thr Thr 115 120 125 Leu Gln Gln Gln Val Ala Thr Glu Ile Glu Thr Ala Pro Thr Leu Ala 130 135 140 Ala Leu Thr Val Arg Lys Leu Arg Phe Ile Asp Lys Val Ser Glu Asp 145 150 155 160 Val Asn Gln Leu Ile Ala Gln Leu Glu Asp 165 170 27 1638 DNA Methylophilus methylotrophus CDS (1)..(1638) 27 atg gca gct aaa gac gta aga ttt ggt gat gac gtt cgc caa aaa atg 48 Met Ala Ala Lys Asp Val Arg Phe Gly Asp Asp Val Arg Gln Lys Met 1 5 10 15 gta aat ggc gtt aac gta ttg gct aac gct gtg cgc gta act ttg ggc 96 Val Asn Gly Val Asn Val Leu Ala Asn Ala Val Arg Val Thr Leu Gly 20 25 30 cct aaa ggc cgt aac gtg gta ttg gag cgt tct ttt ggc gcg cca acc 144 Pro Lys Gly Arg Asn Val Val Leu Glu Arg Ser Phe Gly Ala Pro Thr 35 40 45 atc act aaa gac ggt gtg tct gtg gct aaa gaa atc gaa ttg aaa gac 192 Ile Thr Lys Asp Gly Val Ser Val Ala Lys Glu Ile Glu Leu Lys Asp 50 55 60 aaa ttc gaa aac atg ggc gca cag atg gtg aaa gaa gtg gct tct aaa 240 Lys Phe Glu Asn Met Gly Ala Gln Met Val Lys Glu Val Ala Ser Lys 65 70 75 80 acc aac gac atc gct ggt gac ggt aca acg act gcg act gtg ttg gca 288 Thr Asn Asp Ile Ala Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala 85 90 95 caa gcg atc atc cgc gaa ggc atg aaa tct gtg gct gct ggc atg aac 336 Gln Ala Ile Ile Arg Glu Gly Met Lys Ser Val Ala Ala Gly Met Asn 100 105 110 cca atg gac ctg aag cgc ggt atc gac aaa gcg gtt gaa gcg gcg att 384 Pro Met Asp Leu Lys Arg Gly Ile Asp Lys Ala Val Glu Ala Ala Ile 115 120 125 gct gaa ttg aaa gtg caa tcc aaa ccc tgt acg acc agc aaa gaa atc 432 Ala Glu Leu Lys Val Gln Ser Lys Pro Cys Thr Thr Ser Lys Glu Ile 130 135 140 gcc cag gta ggt tct atc tct gct aac tcc gac act tct gtt ggc caa 480 Ala Gln Val Gly Ser Ile Ser Ala Asn Ser Asp Thr Ser Val Gly Gln 145 150 155 160 att att gct gat gcg atg gac aaa gta ggt aaa gaa ggc gtg atc act 528 Ile Ile Ala Asp Ala Met Asp Lys Val Gly Lys Glu Gly Val Ile Thr 165 170 175 gtt gaa gac ggt tct ggc ttg agc aac gag ctg gac gtg gtt gag ggt 576 Val Glu Asp Gly Ser Gly Leu Ser Asn Glu Leu Asp Val Val Glu Gly 180 185 190 atg caa ttt gat cgc ggt tac ttg tcc cca tac ttc atc aac aac cca 624 Met Gln Phe Asp Arg Gly Tyr Leu Ser Pro Tyr Phe Ile Asn Asn Pro 195 200 205 gag cgc caa att gcg ttg ctg gac aat cct ttt gta ttg ttg cac gac 672 Glu Arg Gln Ile Ala Leu Leu Asp Asn Pro Phe Val Leu Leu His Asp 210 215 220 aag aaa atc tcc aac atc cgt gac ctg ctg cca acc ctg gag caa gtg 720 Lys Lys Ile Ser Asn Ile Arg Asp Leu Leu Pro Thr Leu Glu Gln Val 225 230 235 240 gct aaa gcc ggc cgt cca ttg ctg atc atc gct gaa gat gta gat ggc 768 Ala Lys Ala Gly Arg Pro Leu Leu Ile Ile Ala Glu Asp Val Asp Gly 245 250 255 gaa gct ctg gca acg ttg gta gta aac aac atc cgc ggc atc ctg aaa 816 Glu Ala Leu Ala Thr Leu Val Val Asn Asn Ile Arg Gly Ile Leu Lys 260 265 270 aca acc gct gtg aaa gcg cct ggt ttt ggt gac cgt cgt aaa gcg atg 864 Thr Thr Ala Val Lys Ala Pro Gly Phe Gly Asp Arg Arg Lys Ala Met 275 280 285 ttg gaa gat atc gct gtg ctg acc ggt ggt acc gtg att tct gaa gaa 912 Leu Glu Asp Ile Ala Val Leu Thr Gly Gly Thr Val Ile Ser Glu Glu 290 295 300 gtg ggc ctg aaa ctg gaa ggc gta cag ctg cac gac ctg ggt caa gcc 960 Val Gly Leu Lys Leu Glu Gly Val Gln Leu His Asp Leu Gly Gln Ala 305 310 315 320 aaa cgt atc gaa gtg ggt aaa gaa aac acc atc atc att gat ggc gct 1008 Lys Arg Ile Glu Val Gly Lys Glu Asn Thr Ile Ile Ile Asp Gly Ala 325 330 335 ggt aac gaa gaa gcg atc aaa gcg cgt atc ggc cag atc aaa acc caa 1056 Gly Asn Glu Glu Ala Ile Lys Ala Arg Ile Gly Gln Ile Lys Thr Gln 340 345 350 atc gaa gaa gct tcc agc gac tac gac cgt gaa aaa ctg caa gag cgc 1104 Ile Glu Glu Ala Ser Ser Asp Tyr Asp Arg Glu Lys Leu Gln Glu Arg 355 360 365 gtg gcc aaa ctg gct ggt ggt gtt gca gtg atc aag gtg ggt gct gcg 1152 Val Ala Lys Leu Ala Gly Gly Val Ala Val Ile Lys Val Gly Ala Ala 370 375 380 act gag gtt gaa atg aaa gag aaa aaa gca cgc gtt gaa gat gca ttg 1200 Thr Glu Val Glu Met Lys Glu Lys Lys Ala Arg Val Glu Asp Ala Leu 385 390 395 400 cac gcg act cgc gct gcg gtt gaa gaa ggt atc gtg gcg ggt ggc ggt 1248 His Ala Thr Arg Ala Ala Val Glu Glu Gly Ile Val Ala Gly Gly Gly 405 410 415 gtt gca ttg atc cgt gct cgt gac gcg att gct aaa gtc aaa ggc gaa 1296 Val Ala Leu Ile Arg Ala Arg Asp Ala Ile Ala Lys Val Lys Gly Glu 420 425 430 aac gct gat cag gat gct ggt atc aag atc gtt ctg cgt gcg gtt gaa 1344 Asn Ala Asp Gln Asp Ala Gly Ile Lys Ile Val Leu Arg Ala Val Glu 435 440 445 gaa cca ctg cgc cag atc gtt tct aac gcc ggt gct gaa cca tct gtg 1392 Glu Pro Leu Arg Gln Ile Val Ser Asn Ala Gly Ala Glu Pro Ser Val 450 455 460 gtt gtc agc aac gtt gct gct ggt aaa ggt aac tac ggt tac aac gct 1440 Val Val Ser Asn Val Ala Ala Gly Lys Gly Asn Tyr Gly Tyr Asn Ala 465 470 475 480 gcc aac gaa acc tat ggc gac atg gtt gaa atg ggc gta ctg gat cca 1488 Ala Asn Glu Thr Tyr Gly Asp Met Val Glu Met Gly Val Leu Asp Pro 485 490 495 acc aaa gtg aca cgt tct gcc ttg act aac gca gct tct gtc gct ggc 1536 Thr Lys Val Thr Arg Ser Ala Leu Thr Asn Ala Ala Ser Val Ala Gly 500 505 510 ctg atg ctg acg act gac tgc atg gtc gca gaa ctg cct aaa gaa gat 1584 Leu Met Leu Thr Thr Asp Cys Met Val Ala Glu Leu Pro Lys Glu Asp 515 520 525 gca cct gct gcg ccg gat atg ggc ggg atg ggt ggc atg ggc ggt atg 1632 Ala Pro Ala Ala Pro Asp Met Gly Gly Met Gly Gly Met Gly Gly Met 530 535 540 atg taa 1638 Met 545 28 545 PRT Methylophilus methylotrophus 28 Met Ala Ala Lys Asp Val Arg Phe Gly Asp Asp Val Arg Gln Lys Met 1 5 10 15 Val Asn Gly Val Asn Val Leu Ala Asn Ala Val Arg Val Thr Leu Gly 20 25 30 Pro Lys Gly Arg Asn Val Val Leu Glu Arg Ser Phe Gly Ala Pro Thr 35 40 45 Ile Thr Lys Asp Gly Val Ser Val Ala Lys Glu Ile Glu Leu Lys Asp 50 55 60 Lys Phe Glu Asn Met Gly Ala Gln Met Val Lys Glu Val Ala Ser Lys 65 70 75 80 Thr Asn Asp Ile Ala Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala 85 90 95 Gln Ala Ile Ile Arg Glu Gly Met Lys Ser Val Ala Ala Gly Met Asn 100 105 110 Pro Met Asp Leu Lys Arg Gly Ile Asp Lys Ala Val Glu Ala Ala Ile 115 120 125 Ala Glu Leu Lys Val Gln Ser Lys Pro Cys Thr Thr Ser Lys Glu Ile 130 135 140 Ala Gln Val Gly Ser Ile Ser Ala Asn Ser Asp Thr Ser Val Gly Gln 145 150 155 160 Ile Ile Ala Asp Ala Met Asp Lys Val Gly Lys Glu Gly Val Ile Thr 165 170 175 Val Glu Asp Gly Ser Gly Leu Ser Asn Glu Leu Asp Val Val Glu Gly 180 185 190 Met Gln Phe Asp Arg Gly Tyr Leu Ser Pro Tyr Phe Ile Asn Asn Pro 195 200 205 Glu Arg Gln Ile Ala Leu Leu Asp Asn Pro Phe Val Leu Leu His Asp 210 215 220 Lys Lys Ile Ser Asn Ile Arg Asp Leu Leu Pro Thr Leu Glu Gln Val 225 230 235 240 Ala Lys Ala Gly Arg Pro Leu Leu Ile Ile Ala Glu Asp Val Asp Gly 245 250 255 Glu Ala Leu Ala Thr Leu Val Val Asn Asn Ile Arg Gly Ile Leu Lys 260 265 270 Thr Thr Ala Val Lys Ala Pro Gly Phe Gly Asp Arg Arg Lys Ala Met 275 280 285 Leu Glu Asp Ile Ala Val Leu Thr Gly Gly Thr Val Ile Ser Glu Glu 290 295 300 Val Gly Leu Lys Leu Glu Gly Val Gln Leu His Asp Leu Gly Gln Ala 305 310 315 320 Lys Arg Ile Glu Val Gly Lys Glu Asn Thr Ile Ile Ile Asp Gly Ala 325 330 335 Gly Asn Glu Glu Ala Ile Lys Ala Arg Ile Gly Gln Ile Lys Thr Gln 340 345 350 Ile Glu Glu Ala Ser Ser Asp Tyr Asp Arg Glu Lys Leu Gln Glu Arg 355 360 365 Val Ala Lys Leu Ala Gly Gly Val Ala Val Ile Lys Val Gly Ala Ala 370 375 380 Thr Glu Val Glu Met Lys Glu Lys Lys Ala Arg Val Glu Asp Ala Leu 385 390 395 400 His Ala Thr Arg Ala Ala Val Glu Glu Gly Ile Val Ala Gly Gly Gly 405 410 415 Val Ala Leu Ile Arg Ala Arg Asp Ala Ile Ala Lys Val Lys Gly Glu 420 425 430 Asn Ala Asp Gln Asp Ala Gly Ile Lys Ile Val Leu Arg Ala Val Glu 435 440 445 Glu Pro Leu Arg Gln Ile Val Ser Asn Ala Gly Ala Glu Pro Ser Val 450 455 460 Val Val Ser Asn Val Ala Ala Gly Lys Gly Asn Tyr Gly Tyr Asn Ala 465 470 475 480 Ala Asn Glu Thr Tyr Gly Asp Met Val Glu Met Gly Val Leu Asp Pro 485 490 495 Thr Lys Val Thr Arg Ser Ala Leu Thr Asn Ala Ala Ser Val Ala Gly 500 505 510 Leu Met Leu Thr Thr Asp Cys Met Val Ala Glu Leu Pro Lys Glu Asp 515 520 525 Ala Pro Ala Ala Pro Asp Met Gly Gly Met Gly Gly Met Gly Gly Met 530 535 540 Met 545 29 288 DNA Methylophilus methylotrophus CDS (1)..(288) 29 atg aac att cgt ccg ttg cac gac cgt gta att gtt aaa cgc gct gct 48 Met Asn Ile Arg Pro Leu His Asp Arg Val Ile Val Lys Arg Ala Ala 1 5 10 15 gaa gag cgc acc act gct tct ggc att gtg atc cct gac agt gcg act 96 Glu Glu Arg Thr Thr Ala Ser Gly Ile Val Ile Pro Asp Ser Ala Thr 20 25 30 gag aaa cca gat caa ggt gtg gtt cag gca gtt ggc aat ggc aaa aaa 144 Glu Lys Pro Asp Gln Gly Val Val Gln Ala Val Gly Asn Gly Lys Lys 35 40 45 gat gaa aac ggc aaa gca att gcc ttg gac gtg aaa gtc ggc gac aag 192 Asp Glu Asn Gly Lys Ala Ile Ala Leu Asp Val Lys Val Gly Asp Lys 50 55 60 gta ttg ttc ggc aaa tac tcc ggt caa aca gtg aaa gtc aac ggc gaa 240 Val Leu Phe Gly Lys Tyr Ser Gly Gln Thr Val Lys Val Asn Gly Glu 65 70 75 80 gag ctg ttg gtg atg cgc gaa gaa gac att atg gcg att gtt gag taa 288 Glu Leu Leu Val Met Arg Glu Glu Asp Ile Met Ala Ile Val Glu 85 90 95 30 95 PRT Methylophilus methylotrophus 30 Met Asn Ile Arg Pro Leu His Asp Arg Val Ile Val Lys Arg Ala Ala 1 5 10 15 Glu Glu Arg Thr Thr Ala Ser Gly Ile Val Ile Pro Asp Ser Ala Thr 20 25 30 Glu Lys Pro Asp Gln Gly Val Val Gln Ala Val Gly Asn Gly Lys Lys 35 40 45 Asp Glu Asn Gly Lys Ala Ile Ala Leu Asp Val Lys Val Gly Asp Lys 50 55 60 Val Leu Phe Gly Lys Tyr Ser Gly Gln Thr Val Lys Val Asn Gly Glu 65 70 75 80 Glu Leu Leu Val Met Arg Glu Glu Asp Ile Met Ala Ile Val Glu 85 90 95 31 1620 DNA Methylophilus methylotrophus CDS (1)..(1620) 31 atg gca gca aaa gat gtg aaa ttt cat gat cat gcg cgc act aaa att 48 Met Ala Ala Lys Asp Val Lys Phe His Asp His Ala Arg Thr Lys Ile 1 5 10 15 gta aaa ggc gtg aat atc ctg gcc gat gcg gtc aag gtt acc ttg ggg 96 Val Lys Gly Val Asn Ile Leu Ala Asp Ala Val Lys Val Thr Leu Gly 20 25 30 ccc aaa ggc cgt aat gtc gtg ctg gag cgc agc ttt ggc gcc ccg gtg 144 Pro Lys Gly Arg Asn Val Val Leu Glu Arg Ser Phe Gly Ala Pro Val 35 40 45 att acc aaa gat ggt gtc tcc gtt gcc aag gaa atc gag ttg cag gac 192 Ile Thr Lys Asp Gly Val Ser Val Ala Lys Glu Ile Glu Leu Gln Asp 50 55 60 aag ctg gag aat atg ggc gca caa atg gtg aag caa gtc gct tct aaa 240 Lys Leu Glu Asn Met Gly Ala Gln Met Val Lys Gln Val Ala Ser Lys 65 70 75 80 aca gct gac gtg gcc ggt gac ggt acg act acc gct acc gtg ctg gct 288 Thr Ala Asp Val Ala Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala 85 90 95 cag gcg att gta caa gaa ggg atg aag tca gtc gcc tcc ggc atg aat 336 Gln Ala Ile Val Gln Glu Gly Met Lys Ser Val Ala Ser Gly Met Asn 100 105 110 ccc acc gac cta aaa cgt ggg att gat aaa gcc gtg aca gcc ctg gtg 384 Pro Thr Asp Leu Lys Arg Gly Ile Asp Lys Ala Val Thr Ala Leu Val 115 120 125 gat gag ctt aaa tcc atg tcc aaa gct atc acc acc cat aaa gaa att 432 Asp Glu Leu Lys Ser Met Ser Lys Ala Ile Thr Thr His Lys Glu Ile 130 135 140 gcc caa gtc ggt gcg att tct gcc aac tct gac cat gcc att ggc cag 480 Ala Gln Val Gly Ala Ile Ser Ala Asn Ser Asp His Ala Ile Gly Gln 145 150 155 160 atc atc gcc gat gcc atg gaa aaa gtc ggt aaa gaa ggc gtg att acg 528 Ile Ile Ala Asp Ala Met Glu Lys Val Gly Lys Glu Gly Val Ile Thr 165 170 175 gtg gaa gaa ggt aag tca ctg caa aat gaa ctc gaa gtg gtc gag ggc 576 Val Glu Glu Gly Lys Ser Leu Gln Asn Glu Leu Glu Val Val Glu Gly 180 185 190 atg cag ttt gac cgc ggc tat atc agc ccc tac ttt att aac aac cct 624 Met Gln Phe Asp Arg Gly Tyr Ile Ser Pro Tyr Phe Ile Asn Asn Pro 195 200 205 gac aag cag gta gca gct ctc gat gag ccc atg att ctg ctt tac gac 672 Asp Lys Gln Val Ala Ala Leu Asp Glu Pro Met Ile Leu Leu Tyr Asp 210 215 220 aaa aaa atc agc aat atc cgt gac ttg ttg cca acc ctg gaa aac gtc 720 Lys Lys Ile Ser Asn Ile Arg Asp Leu Leu Pro Thr Leu Glu Asn Val 225 230 235 240 gcc aag gcc aac aaa ccc ttg ctg atc att gcc gaa gac gtg gag ggc 768 Ala Lys Ala Asn Lys Pro Leu Leu Ile Ile Ala Glu Asp Val Glu Gly 245 250 255 gaa gcc ctg gcc acc ctg gtg gtc aat agc atg cgc ggt atc ctc aaa 816 Glu Ala Leu Ala Thr Leu Val Val Asn Ser Met Arg Gly Ile Leu Lys 260 265 270 gtg gtt gct gtg aaa gca cct ggc ttt ggt gac cgc cgc aaa gcc atg 864 Val Val Ala Val Lys Ala Pro Gly Phe Gly Asp Arg Arg Lys Ala Met 275 280 285 ctg gaa gac att gcc gtc ctg acc ggg gcc act gtg gtt tcc gaa gaa 912 Leu Glu Asp Ile Ala Val Leu Thr Gly Ala Thr Val Val Ser Glu Glu 290 295 300 acc ggc atg caa ctg gaa aaa gtg act ctt gaa cac ttg ggc cac gcc 960 Thr Gly Met Gln Leu Glu Lys Val Thr Leu Glu His Leu Gly His Ala 305 310 315 320 aag cgc gtt gaa gtg caa aaa gag aat acc atc att att gat ggc gca 1008 Lys Arg Val Glu Val Gln Lys Glu Asn Thr Ile Ile Ile Asp Gly Ala 325 330 335 ggt gat gca gcc aag att aag gcc cgc gtg cag tcc atc cgg act caa 1056 Gly Asp Ala Ala Lys Ile Lys Ala Arg Val Gln Ser Ile Arg Thr Gln 340 345 350 ata gag gaa gcc acc tct gat tat gac aag gaa aaa ctg cag gag cgc 1104 Ile Glu Glu Ala Thr Ser Asp Tyr Asp Lys Glu Lys Leu Gln Glu Arg 355 360 365 gtg gcc aaa ctg ggt ggt ggc gtg gcg gtg att

aaa atc ggc gcc gcg 1152 Val Ala Lys Leu Gly Gly Gly Val Ala Val Ile Lys Ile Gly Ala Ala 370 375 380 acc gaa gtc gaa atg aaa gag aaa aaa gac cgt gtc gat gat gct cta 1200 Thr Glu Val Glu Met Lys Glu Lys Lys Asp Arg Val Asp Asp Ala Leu 385 390 395 400 cac gcc aca cgg gct gcg gtg gaa gaa ggc atc gtt cca ggt ggc ggt 1248 His Ala Thr Arg Ala Ala Val Glu Glu Gly Ile Val Pro Gly Gly Gly 405 410 415 gtg gct ctg ctg cgt gcc cgc agc cgc atg agc act ctc aag ggc gat 1296 Val Ala Leu Leu Arg Ala Arg Ser Arg Met Ser Thr Leu Lys Gly Asp 420 425 430 aat gat gac cag gaa gcc ggt atc cgc att gtg ttg cgt gca ata gaa 1344 Asn Asp Asp Gln Glu Ala Gly Ile Arg Ile Val Leu Arg Ala Ile Glu 435 440 445 gag ccc tta cgc gcg att gtg aaa aat gca ggt gaa gag cct tct gtc 1392 Glu Pro Leu Arg Ala Ile Val Lys Asn Ala Gly Glu Glu Pro Ser Val 450 455 460 gtc atc gcc aag gtg ctt gag gca aca ggt aat acc ggc tac aac gca 1440 Val Ile Ala Lys Val Leu Glu Ala Thr Gly Asn Thr Gly Tyr Asn Ala 465 470 475 480 gcc acc ggt gaa tat gtg gat atg gtg gaa acc ggt gtg gtg gac cca 1488 Ala Thr Gly Glu Tyr Val Asp Met Val Glu Thr Gly Val Val Asp Pro 485 490 495 acc aag gtg acc cgc act gcc ctg caa aat gca gcc tcg att gcc ggt 1536 Thr Lys Val Thr Arg Thr Ala Leu Gln Asn Ala Ala Ser Ile Ala Gly 500 505 510 ctg att tta acc acc gac gcc acc gtg gct gaa cta ccc aaa gag gag 1584 Leu Ile Leu Thr Thr Asp Ala Thr Val Ala Glu Leu Pro Lys Glu Glu 515 520 525 aaa aag gcg cca gcc atg cca gaa atg gag tac taa 1620 Lys Lys Ala Pro Ala Met Pro Glu Met Glu Tyr 530 535 32 539 PRT Methylophilus methylotrophus 32 Met Ala Ala Lys Asp Val Lys Phe His Asp His Ala Arg Thr Lys Ile 1 5 10 15 Val Lys Gly Val Asn Ile Leu Ala Asp Ala Val Lys Val Thr Leu Gly 20 25 30 Pro Lys Gly Arg Asn Val Val Leu Glu Arg Ser Phe Gly Ala Pro Val 35 40 45 Ile Thr Lys Asp Gly Val Ser Val Ala Lys Glu Ile Glu Leu Gln Asp 50 55 60 Lys Leu Glu Asn Met Gly Ala Gln Met Val Lys Gln Val Ala Ser Lys 65 70 75 80 Thr Ala Asp Val Ala Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala 85 90 95 Gln Ala Ile Val Gln Glu Gly Met Lys Ser Val Ala Ser Gly Met Asn 100 105 110 Pro Thr Asp Leu Lys Arg Gly Ile Asp Lys Ala Val Thr Ala Leu Val 115 120 125 Asp Glu Leu Lys Ser Met Ser Lys Ala Ile Thr Thr His Lys Glu Ile 130 135 140 Ala Gln Val Gly Ala Ile Ser Ala Asn Ser Asp His Ala Ile Gly Gln 145 150 155 160 Ile Ile Ala Asp Ala Met Glu Lys Val Gly Lys Glu Gly Val Ile Thr 165 170 175 Val Glu Glu Gly Lys Ser Leu Gln Asn Glu Leu Glu Val Val Glu Gly 180 185 190 Met Gln Phe Asp Arg Gly Tyr Ile Ser Pro Tyr Phe Ile Asn Asn Pro 195 200 205 Asp Lys Gln Val Ala Ala Leu Asp Glu Pro Met Ile Leu Leu Tyr Asp 210 215 220 Lys Lys Ile Ser Asn Ile Arg Asp Leu Leu Pro Thr Leu Glu Asn Val 225 230 235 240 Ala Lys Ala Asn Lys Pro Leu Leu Ile Ile Ala Glu Asp Val Glu Gly 245 250 255 Glu Ala Leu Ala Thr Leu Val Val Asn Ser Met Arg Gly Ile Leu Lys 260 265 270 Val Val Ala Val Lys Ala Pro Gly Phe Gly Asp Arg Arg Lys Ala Met 275 280 285 Leu Glu Asp Ile Ala Val Leu Thr Gly Ala Thr Val Val Ser Glu Glu 290 295 300 Thr Gly Met Gln Leu Glu Lys Val Thr Leu Glu His Leu Gly His Ala 305 310 315 320 Lys Arg Val Glu Val Gln Lys Glu Asn Thr Ile Ile Ile Asp Gly Ala 325 330 335 Gly Asp Ala Ala Lys Ile Lys Ala Arg Val Gln Ser Ile Arg Thr Gln 340 345 350 Ile Glu Glu Ala Thr Ser Asp Tyr Asp Lys Glu Lys Leu Gln Glu Arg 355 360 365 Val Ala Lys Leu Gly Gly Gly Val Ala Val Ile Lys Ile Gly Ala Ala 370 375 380 Thr Glu Val Glu Met Lys Glu Lys Lys Asp Arg Val Asp Asp Ala Leu 385 390 395 400 His Ala Thr Arg Ala Ala Val Glu Glu Gly Ile Val Pro Gly Gly Gly 405 410 415 Val Ala Leu Leu Arg Ala Arg Ser Arg Met Ser Thr Leu Lys Gly Asp 420 425 430 Asn Asp Asp Gln Glu Ala Gly Ile Arg Ile Val Leu Arg Ala Ile Glu 435 440 445 Glu Pro Leu Arg Ala Ile Val Lys Asn Ala Gly Glu Glu Pro Ser Val 450 455 460 Val Ile Ala Lys Val Leu Glu Ala Thr Gly Asn Thr Gly Tyr Asn Ala 465 470 475 480 Ala Thr Gly Glu Tyr Val Asp Met Val Glu Thr Gly Val Val Asp Pro 485 490 495 Thr Lys Val Thr Arg Thr Ala Leu Gln Asn Ala Ala Ser Ile Ala Gly 500 505 510 Leu Ile Leu Thr Thr Asp Ala Thr Val Ala Glu Leu Pro Lys Glu Glu 515 520 525 Lys Lys Ala Pro Ala Met Pro Glu Met Glu Tyr 530 535 33 318 DNA Methylophilus methylotrophus CDS (1)..(318) 33 atg aat att cgt ccc ctg tat gac cgc gtc att gtc aag cga gtc gag 48 Met Asn Ile Arg Pro Leu Tyr Asp Arg Val Ile Val Lys Arg Val Glu 1 5 10 15 caa caa cgg acg acc gct tcc ggc att gtg atc cct gac act gca gca 96 Gln Gln Arg Thr Thr Ala Ser Gly Ile Val Ile Pro Asp Thr Ala Ala 20 25 30 gaa aag cca gag caa gga gag gtc att gcc gtg ggc agt ggc aaa cag 144 Glu Lys Pro Glu Gln Gly Glu Val Ile Ala Val Gly Ser Gly Lys Gln 35 40 45 ctg cag gac ggt agc ctg cga cca ctg gaa gtc aag gtg ggc aac cat 192 Leu Gln Asp Gly Ser Leu Arg Pro Leu Glu Val Lys Val Gly Asn His 50 55 60 atc ctg ttt ggc aag tat tct ggt caa acg gtc aag ctt aac ggc gag 240 Ile Leu Phe Gly Lys Tyr Ser Gly Gln Thr Val Lys Leu Asn Gly Glu 65 70 75 80 gaa ctg ctg gtc atg cgt gag gaa gac att ctc ggc gtg att gaa ccc 288 Glu Leu Leu Val Met Arg Glu Glu Asp Ile Leu Gly Val Ile Glu Pro 85 90 95 agt cct gcc gac ctc aaa aaa gtc gct taa 318 Ser Pro Ala Asp Leu Lys Lys Val Ala 100 105 34 105 PRT Methylophilus methylotrophus 34 Met Asn Ile Arg Pro Leu Tyr Asp Arg Val Ile Val Lys Arg Val Glu 1 5 10 15 Gln Gln Arg Thr Thr Ala Ser Gly Ile Val Ile Pro Asp Thr Ala Ala 20 25 30 Glu Lys Pro Glu Gln Gly Glu Val Ile Ala Val Gly Ser Gly Lys Gln 35 40 45 Leu Gln Asp Gly Ser Leu Arg Pro Leu Glu Val Lys Val Gly Asn His 50 55 60 Ile Leu Phe Gly Lys Tyr Ser Gly Gln Thr Val Lys Leu Asn Gly Glu 65 70 75 80 Glu Leu Leu Val Met Arg Glu Glu Asp Ile Leu Gly Val Ile Glu Pro 85 90 95 Ser Pro Ala Asp Leu Lys Lys Val Ala 100 105

Claims

1. An isolated polynucleotide which encodes a protein comprising the amino acid sequence of SEQ ID NO:18.

2. A vector comprising the isolated polynucleotide of claim 1.

3. A host cell comprising the isolated polynucleotide of claim 1.

4. The host cell of claim 3, which is a Methylophilus bacterium.

5. The host cell of claim 4, which is a Methylophilus methylotrophus bacterium.

6. A method of producing at least one amino acid comprising culturing the host cell of claim 3 for a time and under conditions suitable for producing the amino acid; and collecting the amino acid produced.

7. The method of claim 6, wherein said at least one amino acid is an L-amino acid.

8. The method of claim 6, wherein said host cell is a Methylophilus bacterium.

9. The method of claim 6, wherein said host cell is a Methylophilus methylotrophus bacterium.

10. An isolated polynucleotide comprising the nucleotide sequence of SEQ ID NO: 17.

11. A vector comprising the isolated polynucleotide of claim 10.

12. A host cell comprising the isolated polynucleotide of claim 10.

13. The host cell of claim 12, which is a Methylophilus bacterium.

14. The host cell of claim 12, which is a Methylophilus methylotrophus bacterium.

15. A method of producing at least one amino acid comprising culturing the host cell of claim 12 for a time and under conditions suitable for producing the amino acid; and collecting the amino acid produced.

16. The method of claim 15, wherein said at least one amino acid is an L-amino acid.

17. The method of claim 15, wherein said host cell is a Methylophilus bacterium.

18. The method of claim 15, wherein said host cell is a Methylophilus methylotrophus bacterium.

19. An isolated polynucleotide, which hybridizes under stringent conditions to the isolated polynucleotides of claim 10.

20. A vector comprising the isolated polynucleotide of claim 19.

21. A host cell comprising the isolated polynucleotide of claim 19.

22. A method of producing at least one amino acid comprising culturing the host cell of claim 21 for a time and under conditions suitable for producing the amino acid; and collecting the amino acid produced.

23. The method of claim 22, wherein said at least one amino acid is an L-amino acid.

24. The method of claim 22, wherein said host cell is a Methylophilus bacterium.

25. The method of claim 22, wherein said host cell is a Methylophilus methylotrophus bacterium.

26. An isolated polynucleotide, which is at least 95% identical to the polynucleotide of claim 10.

27. A vector comprising the isolated polynucleotide of claim 26.

28. A host cell comprising the isolated polynucleotide of claim 26.

29. A method of producing at least one amino acid comprising culturing the host cell of claim 28 for a time and under conditions suitable for producing the amino acid; and collecting the amino acid produced.

30. The method of claim 29, wherein said at least one amino acid is an L-amino acid.

31. The method of claim 29, wherein said host cell is a Methylophilus bacterium.

32. The method of claim 29, wherein said host cell is a Methylophilus methylotrophus bacterium.