The complete genome sequence of Escherichia coli K-12

Science

Volumn 277, Issue 5331, 1997, Pages 1453-1462

  Blattner, Frederick R ^a   Plunkett III, Guy ^a   Bloch, Craig A ^b   Perna, Nicole T ^a   Burland, Valerie ^c   Riley, Monica ^d   Collado Vides, Julio ^e   Glasner, Jeremy D ^a   Rode, Christopher K ^b   Mayhew, George F ^a   Gregor, Jason ^a   Davis, Nelson Wayne ^a   Kirkpatrick, Heather A ^a   Goeden, Michael A ^a   Rose, Debra J ^a   Mau, Bob ^a   Shao, Ying ^a  

^a UNIVERSITY OF WISCONSIN MADISON   (United States)

^b UNIVERSITY OF MICHIGAN MEDICAL SCHOOL   (United States)

^c FMC CORPORATION   (United States)

^d MARINE BIOLOGICAL LABORATORY   (United States)

^e INSTITUTO DE CIENCIAS FÍSICAS   (Mexico)

Author keywords

[No Author keywords available]

Indexed keywords

ABC TRANSPORTER; BACTERIAL DNA; BACTERIAL PROTEIN;

BACTERIAL GENETICS; BASE PAIRING; DNA REPLICATION; DNA SEQUENCE; ESCHERICHIA COLI; GENE MAPPING; GENOME; NONHUMAN; POLYMERASE CHAIN REACTION; PRIORITY JOURNAL; REVIEW;

EID: 15444350252     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5331.1453     Document Type: Review

References (109)

1. F. R. Blattner, Science 222, 719 (1983). Escherichia coli has been the subject of extensive monographs, the most recent of which is (2).
2. Escherichia coli and Salmonella Cellular and Molecular Biology, F. C. Neidhardt et al., Eds. (ASM Press, Washington, DC, 1996).
3. The publicly available complete genome sequences are those of Haemophilus influenzae Rd [R. D. Fleischmann et al., Science 269, 496 (1995)], Mycoplasma genitalium [C. M. Fraser et al., ibid. 270, 397 (1995)], Methanococcus jannaschii [C. J. Bult et al., ibid. 273, 1058 (1996)], Mycoplasma pneumoniae [R. Himmelreich et al., Nucleic Acids Res. 24, 4420 (1996)], Synechocystis sp. strain PCC6803 [T. Kaneko et al., DNA Res. 3,109 (1996)], and Saccharomyces cerevisiae [A. Goffeau et al., Science 274, 546 (1996)].
4. The publicly available complete genome sequences are those of Haemophilus influenzae Rd [R. D. Fleischmann et al., Science 269, 496 (1995)], Mycoplasma genitalium [C. M. Fraser et al., ibid. 270, 397 (1995)], Methanococcus jannaschii [C. J. Bult et al., ibid. 273, 1058 (1996)], Mycoplasma pneumoniae [R. Himmelreich et al., Nucleic Acids Res. 24, 4420 (1996)], Synechocystis sp. strain PCC6803 [T. Kaneko et al., DNA Res. 3,109 (1996)], and Saccharomyces cerevisiae [A. Goffeau et al., Science 274, 546 (1996)].
5. The publicly available complete genome sequences are those of Haemophilus influenzae Rd [R. D. Fleischmann et al., Science 269, 496 (1995)], Mycoplasma genitalium [C. M. Fraser et al., ibid. 270, 397 (1995)], Methanococcus jannaschii [C. J. Bult et al., ibid. 273, 1058 (1996)], Mycoplasma pneumoniae [R. Himmelreich et al., Nucleic Acids Res. 24, 4420 (1996)], Synechocystis sp. strain PCC6803 [T. Kaneko et al., DNA Res. 3,109 (1996)], and Saccharomyces cerevisiae [A. Goffeau et al., Science 274, 546 (1996)].
6. The publicly available complete genome sequences are those of Haemophilus influenzae Rd [R. D. Fleischmann et al., Science 269, 496 (1995)], Mycoplasma genitalium [C. M. Fraser et al., ibid. 270, 397 (1995)], Methanococcus jannaschii [C. J. Bult et al., ibid. 273, 1058 (1996)], Mycoplasma pneumoniae [R. Himmelreich et al., Nucleic Acids Res. 24, 4420 (1996)], Synechocystis sp. strain PCC6803 [T. Kaneko et al., DNA Res. 3,109 (1996)], and Saccharomyces cerevisiae [A. Goffeau et al., Science 274, 546 (1996)].
7. The publicly available complete genome sequences are those of Haemophilus influenzae Rd [R. D. Fleischmann et al., Science 269, 496 (1995)], Mycoplasma genitalium [C. M. Fraser et al., ibid. 270, 397 (1995)], Methanococcus jannaschii [C. J. Bult et al., ibid. 273, 1058 (1996)], Mycoplasma pneumoniae [R. Himmelreich et al., Nucleic Acids Res. 24, 4420 (1996)], Synechocystis sp. strain PCC6803 [T. Kaneko et al., DNA Res. 3,109 (1996)], and Saccharomyces cerevisiae [A. Goffeau et al., Science 274, 546 (1996)].
8. The publicly available complete genome sequences are those of Haemophilus influenzae Rd [R. D. Fleischmann et al., Science 269, 496 (1995)], Mycoplasma genitalium [C. M. Fraser et al., ibid. 270, 397 (1995)], Methanococcus jannaschii [C. J. Bult et al., ibid. 273, 1058 (1996)], Mycoplasma pneumoniae [R. Himmelreich et al., Nucleic Acids Res. 24, 4420 (1996)], Synechocystis sp. strain PCC6803 [T. Kaneko et al., DNA Res. 3,109 (1996)], and Saccharomyces cerevisiae [A. Goffeau et al., Science 274, 546 (1996)].
9. S.-E. Chuang, D. L. Daniels, F. R. Blattner, J. Bacteriol. 175, 2026 (1993); D. J. Lockart et al., Nature Biotechnol. 14, 1675 (1996).
10. S.-E. Chuang, D. L. Daniels, F. R. Blattner, J. Bacteriol. 175, 2026 (1993); D. J. Lockart et al., Nature Biotechnol. 14, 1675 (1996).
11. M. Riley and B. Labedan, J. Mol. Biol. 269, 1 (1997).
12. F. C. Neidhardt, in (2), vol. 2, pp. 1-3.
13. B. Bachmann, in (2), vol. 2, pp. 2460-2488.
14. K. F. Jensen, J. Bacteriol. 175, 3401 (1993).
15. R. P. Lawther et al., ibid. 149, 294 (1982).
16. D. Liu and P. R. Reeves, Microbiology 140, 49 (1994).
17. T. Yura et al., Nucleic Acids Res. 20, 3305 (1992); N. Fujita, H. Mori, T. Yura, A. Ishihama, ibid. 22, 1637 (1994); T. Oshima et al., DNA Res. 3, 137 (1996); H. Aiba et al., ibid., p. 363; T. Itoh et al., ibid., p. 379.
18. T. Yura et al., Nucleic Acids Res. 20, 3305 (1992); N. Fujita, H. Mori, T. Yura, A. Ishihama, ibid. 22, 1637 (1994); T. Oshima et al., DNA Res. 3, 137 (1996); H. Aiba et al., ibid., p. 363; T. Itoh et al., ibid., p. 379.
19. T. Yura et al., Nucleic Acids Res. 20, 3305 (1992); N. Fujita, H. Mori, T. Yura, A. Ishihama, ibid. 22, 1637 (1994); T. Oshima et al., DNA Res. 3, 137 (1996); H. Aiba et al., ibid., p. 363; T. Itoh et al., ibid., p. 379.
20. T. Yura et al., Nucleic Acids Res. 20, 3305 (1992); N. Fujita, H. Mori, T. Yura, A. Ishihama, ibid. 22, 1637 (1994); T. Oshima et al., DNA Res. 3, 137 (1996); H. Aiba et al., ibid., p. 363; T. Itoh et al., ibid., p. 379.
21. T. Yura et al., Nucleic Acids Res. 20, 3305 (1992); N. Fujita, H. Mori, T. Yura, A. Ishihama, ibid. 22, 1637 (1994); T. Oshima et al., DNA Res. 3, 137 (1996); H. Aiba et al., ibid., p. 363; T. Itoh et al., ibid., p. 379.
22. V. Burland, D. L. Daniels, G. Plunkett III, F. R. Blattner, Nucleic Acids Res. 21, 3385 (1993).
23. Six segments of the genome were sequenced using radioactive chemistry (14) [D. L. Daniels, G. Plunkett III, V. Burland, F. R. Blattner, Science 257, 771 (1992); G. Plunkett III, V. Burland, D. L. Daniels, F. R. Blattner, Nucleic Acids Res. 21, 3391 (1993); F. R. Blattner, V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, ibid., p. 5408; H. J. Sofia, V. Burland, D. L Daniels, G. Plunkett III, F. R. Blattner, ibid. 22, 2576 (1994); V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, F. R. Blattner, ibid. 23, 2105 (1995)]. We determined experimentally that deoxyinosine tríphosphate (dITP) is the most effective analog for resolving G-C compressions, although it also causes premature termination. With radioactive sequencing, a dITP sequence lane must be run in addition to, rather than in place of, a deoxyguanosine triphosphate (dGTP) run. For efficiency in the areas of E. coli we sequenced radioactively, tiling software was used to select a minimal set of M13 clones for resequencing with dITP after the bulk of the assembly had been completed with dGTP. On the other hand, because prematurely terminated chains are not labeled by the fluorophore with dye-terminator fluorescent sequencing, dITP can substitute totally for dGTP and can be used for all routine data collection.
24. Six segments of the genome were sequenced using radioactive chemistry (14) [D. L. Daniels, G. Plunkett III, V. Burland, F. R. Blattner, Science 257, 771 (1992); G. Plunkett III, V. Burland, D. L. Daniels, F. R. Blattner, Nucleic Acids Res. 21, 3391 (1993); F. R. Blattner, V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, ibid., p. 5408; H. J. Sofia, V. Burland, D. L Daniels, G. Plunkett III, F. R. Blattner, ibid. 22, 2576 (1994); V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, F. R. Blattner, ibid. 23, 2105 (1995)]. We determined experimentally that deoxyinosine tríphosphate (dITP) is the most effective analog for resolving G-C compressions, although it also causes premature termination. With radioactive sequencing, a dITP sequence lane must be run in addition to, rather than in place of, a deoxyguanosine triphosphate (dGTP) run. For efficiency in the areas of E. coli we sequenced radioactively, tiling software was used to select a minimal set of M13 clones for resequencing with dITP after the bulk of the assembly had been completed with dGTP. On the other hand, because prematurely terminated chains are not labeled by the fluorophore with dye-terminator fluorescent sequencing, dITP can substitute totally for dGTP and can be used for all routine data collection.
25. Six segments of the genome were sequenced using radioactive chemistry (14) [D. L. Daniels, G. Plunkett III, V. Burland, F. R. Blattner, Science 257, 771 (1992); G. Plunkett III, V. Burland, D. L. Daniels, F. R. Blattner, Nucleic Acids Res. 21, 3391 (1993); F. R. Blattner, V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, ibid., p. 5408; H. J. Sofia, V. Burland, D. L Daniels, G. Plunkett III, F. R. Blattner, ibid. 22, 2576 (1994); V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, F. R. Blattner, ibid. 23, 2105 (1995)]. We determined experimentally that deoxyinosine tríphosphate (dITP) is the most effective analog for resolving G-C compressions, although it also causes premature termination. With radioactive sequencing, a dITP sequence lane must be run in addition to, rather than in place of, a deoxyguanosine triphosphate (dGTP) run. For efficiency in the areas of E. coli we sequenced radioactively, tiling software was used to select a minimal set of M13 clones for resequencing with dITP after the bulk of the assembly had been completed with dGTP. On the other hand, because prematurely terminated chains are not labeled by the fluorophore with dye-terminator fluorescent sequencing, dITP can substitute totally for dGTP and can be used for all routine data collection.
26. Six segments of the genome were sequenced using radioactive chemistry (14) [D. L. Daniels, G. Plunkett III, V. Burland, F. R. Blattner, Science 257, 771 (1992); G. Plunkett III, V. Burland, D. L. Daniels, F. R. Blattner, Nucleic Acids Res. 21, 3391 (1993); F. R. Blattner, V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, ibid., p. 5408; H. J. Sofia, V. Burland, D. L Daniels, G. Plunkett III, F. R. Blattner, ibid. 22, 2576 (1994); V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, F. R. Blattner, ibid. 23, 2105 (1995)]. We determined experimentally that deoxyinosine tríphosphate (dITP) is the most effective analog for resolving G-C compressions, although it also causes premature termination. With radioactive sequencing, a dITP sequence lane must be run in addition to, rather than in place of, a deoxyguanosine triphosphate (dGTP) run. For efficiency in the areas of E. coli we sequenced radioactively, tiling software was used to select a minimal set of M13 clones for resequencing with dITP after the bulk of the assembly had been completed with dGTP. On the other hand, because prematurely terminated chains are not labeled by the fluorophore with dye-terminator fluorescent sequencing, dITP can substitute totally for dGTP and can be used for all routine data collection.
27. Six segments of the genome were sequenced using radioactive chemistry (14) [D. L. Daniels, G. Plunkett III, V. Burland, F. R. Blattner, Science 257, 771 (1992); G. Plunkett III, V. Burland, D. L. Daniels, F. R. Blattner, Nucleic Acids Res. 21, 3391 (1993); F. R. Blattner, V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, ibid., p. 5408; H. J. Sofia, V. Burland, D. L Daniels, G. Plunkett III, F. R. Blattner, ibid. 22, 2576 (1994); V. Burland, G. Plunkett III, H. J. Sofia, D. L. Daniels, F. R. Blattner, ibid. 23, 2105 (1995)]. We determined experimentally that deoxyinosine tríphosphate (dITP) is the most effective analog for resolving G-C compressions, although it also causes premature termination. With radioactive sequencing, a dITP sequence lane must be run in addition to, rather than in place of, a deoxyguanosine triphosphate (dGTP) run. For efficiency in the areas of E. coli we sequenced radioactively, tiling software was used to select a minimal set of M13 clones for resequencing with dITP after the bulk of the assembly had been completed with dGTP. On the other hand, because prematurely terminated chains are not labeled by the fluorophore with dye-terminator fluorescent sequencing, dITP can substitute totally for dGTP and can be used for all routine data collection.
28. V. Burland, G. Plunkett III, D. L. Daniels, F. R. Blattner, Genomics 16, 551 (1993).
29. D. L. Daniels, in The Bacterial Chromosome, K. Drlica and M. Riley, Eds. (American Society for Microbiology, Washington, DC, 1990), pp. 43-51. It was often necessary to resequence overlapping regions between adjacent clones, and screening to remove lambda vector sequences before sequencing was costly. Occasionally we found deleted, mismapped, or chimeric lambda clones that created unexpected gaps in genome coverage.
30. Although the 1-μg yield of popout plasmid [G. Pósfai et al., Nucleic Acids Res. 22, 2392 (1994)] was low for early shotgun protocols, the assemblies were successful when supplemented with lambda clone and long-range PCR data. The main problem with extending this approach was the need to specifically engineer each popout plasmid by insertional recombination into the host.
31. I-Sce I is a site-specific intron-encoded homing endonuclease from yeast [A. Perrin, M. Buckle, B. Dujon, EMBO J. 12, 2939 (1993)], whose 18-bp non-palindromic recognition site is absent from E. coli (C. A. Bloch and C. K. Rode, unpublished data). Single I-Sce I sites were introduced into MG1655 on a transposable element to produce a mapped collection of strains, each with a unique I-Sce I site [C. K. Rode, V. H. Obreque, C. A. Bloch, Gene 166, 1 (1995); C. A. Bloch, C. K. Rode, V. H. Obreque, J. Mahillon, Biochem. Biophys. Res. Commun. 223, 104 (1996)]. P1 transduction was used to combine sites in pairs, permitting isolation of I-Sce I fragments as single bands by pulsed-field gel electrophoresis. Sequencing confirmed the expected nine-base overlap between adjacent fragments. Although the background contamination for entire I-Sce I fragment shotguns ranged from 15 to 30%, we occasionally observed individual preparative gels that seemed to have <5% background, as assessed from gel images. We therefore suspect that improvements in gel handling and electrophoretic conditions could improve the overall quality of the fragment preparations.
32. I-Sce I is a site-specific intron-encoded homing endonuclease from yeast [A. Perrin, M. Buckle, B. Dujon, EMBO J. 12, 2939 (1993)], whose 18-bp non-palindromic recognition site is absent from E. coli (C. A. Bloch and C. K. Rode, unpublished data). Single I-Sce I sites were introduced into MG1655 on a transposable element to produce a mapped collection of strains, each with a unique I-Sce I site [C. K. Rode, V. H. Obreque, C. A. Bloch, Gene 166, 1 (1995); C. A. Bloch, C. K. Rode, V. H. Obreque, J. Mahillon, Biochem. Biophys. Res. Commun. 223, 104 (1996)]. P1 transduction was used to combine sites in pairs, permitting isolation of I-Sce I fragments as single bands by pulsed-field gel electrophoresis. Sequencing confirmed the expected nine-base overlap between adjacent fragments. Although the background contamination for entire I-Sce I fragment shotguns ranged from 15 to 30%, we occasionally observed individual preparative gels that seemed to have <5% background, as assessed from gel images. We therefore suspect that improvements in gel handling and electrophoretic conditions could improve the overall quality of the fragment preparations.
33. I-Sce I is a site-specific intron-encoded homing endonuclease from yeast [A. Perrin, M. Buckle, B. Dujon, EMBO J. 12, 2939 (1993)], whose 18-bp non-palindromic recognition site is absent from E. coli (C. A. Bloch and C. K. Rode, unpublished data). Single I-Sce I sites were introduced into MG1655 on a transposable element to produce a mapped collection of strains, each with a unique I-Sce I site [C. K. Rode, V. H. Obreque, C. A. Bloch, Gene 166, 1 (1995); C. A. Bloch, C. K. Rode, V. H. Obreque, J. Mahillon, Biochem. Biophys. Res. Commun. 223, 104 (1996)]. P1 transduction was used to combine sites in pairs, permitting isolation of I-Sce I fragments as single bands by pulsed-field gel electrophoresis. Sequencing confirmed the expected nine-base overlap between adjacent fragments. Although the background contamination for entire I-Sce I fragment shotguns ranged from 15 to 30%, we occasionally observed individual preparative gels that seemed to have <5% background, as assessed from gel images. We therefore suspect that improvements in gel handling and electrophoretic conditions could improve the overall quality of the fragment preparations.
34. I-Sce I is a site-specific intron-encoded homing endonuclease from yeast [A. Perrin, M. Buckle, B. Dujon, EMBO J. 12, 2939 (1993)], whose 18-bp non-palindromic recognition site is absent from E. coli (C. A. Bloch and C. K. Rode, unpublished data). Single I-Sce I sites were introduced into MG1655 on a transposable element to produce a mapped collection of strains, each with a unique I-Sce I site [C. K. Rode, V. H. Obreque, C. A. Bloch, Gene 166, 1 (1995); C. A. Bloch, C. K. Rode, V. H. Obreque, J. Mahillon, Biochem. Biophys. Res. Commun. 223, 104 (1996)]. P1 transduction was used to combine sites in pairs, permitting isolation of I-Sce I fragments as single bands by pulsed-field gel electrophoresis. Sequencing confirmed the expected nine-base overlap between adjacent fragments. Although the background contamination for entire I-Sce I fragment shotguns ranged from 15 to 30%, we occasionally observed individual preparative gels that seemed to have <5% background, as assessed from gel images. We therefore suspect that improvements in gel handling and electrophoretic conditions could improve the overall quality of the fragment preparations.
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37. Codon usage statistics [M. Borodovsky and J. McIninch, Comput. Chem. 17, 123 (1993); M. Gribskov, J. Devereux, R. R. Burgess, Nucleic Acids Res. 12, 539 (1984)] were graphically displayed by means of the program Geneplot (DNASTAR). Protein searches were to SWISS-PROT release 34 [A. Bairoch and R. Apweiler, ibid. 24, 21 (1996)]. The Link database is described in A. J. Link, thesis, Harvard University (1994). Signal peptide searches used an unpublished BASIC program written by F. R. B. Predictions for ribosomal binding sites were provided by W. S. Hayes and M. Borodovsky (personal communication).
38. Codon usage statistics [M. Borodovsky and J. McIninch, Comput. Chem. 17, 123 (1993); M. Gribskov, J. Devereux, R. R. Burgess, Nucleic Acids Res. 12, 539 (1984)] were graphically displayed by means of the program Geneplot (DNASTAR). Protein searches were to SWISS-PROT release 34 [A. Bairoch and R. Apweiler, ibid. 24, 21 (1996)]. The Link database is described in A. J. Link, thesis, Harvard University (1994). Signal peptide searches used an unpublished BASIC program written by F. R. B. Predictions for ribosomal binding sites were provided by W. S. Hayes and M. Borodovsky (personal communication).
39. Codon usage statistics [M. Borodovsky and J. McIninch, Comput. Chem. 17, 123 (1993); M. Gribskov, J. Devereux, R. R. Burgess, Nucleic Acids Res. 12, 539 (1984)] were graphically displayed by means of the program Geneplot (DNASTAR). Protein searches were to SWISS-PROT release 34 [A. Bairoch and R. Apweiler, ibid. 24, 21 (1996)]. The Link database is described in A. J. Link, thesis, Harvard University (1994). Signal peptide searches used an unpublished BASIC program written by F. R. B. Predictions for ribosomal binding sites were provided by W. S. Hayes and M. Borodovsky (personal communication).
40. Codon usage statistics [M. Borodovsky and J. McIninch, Comput. Chem. 17, 123 (1993); M. Gribskov, J. Devereux, R. R. Burgess, Nucleic Acids Res. 12, 539 (1984)] were graphically displayed by means of the program Geneplot (DNASTAR). Protein searches were to SWISS-PROT release 34 [A. Bairoch and R. Apweiler, ibid. 24, 21 (1996)]. The Link database is described in A. J. Link, thesis, Harvard University (1994). Signal peptide searches used an unpublished BASIC program written by F. R. B. Predictions for ribosomal binding sites were provided by W. S. Hayes and M. Borodovsky (personal communication).
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44. Similarity searches were conducted using both the DeCypher II hardware-software system (Time Logic Inc., Incline Village, NV) and the PepPepSearch program of the Darwin suite at Zurich, http:// cbrg.inf.ethz.ch/ [G. H. Gonnet, M. A. Cohen, S. A. Benner, Science 256, 1443 (1992)]. PepPepSearch returns up to 30 hit sequences per query, and returns each pairwise alignment and the corresponding PAM scores. For most of the cases, only matches with PAM < 200 were used. See B. Labedan and M. Riley, Mol. Biol. Evol. 12, 980 (1995).
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105. P22 [D. F. Lindsey, C. Martinez, J. R. Walker, J. Bacteriol. 174, 3834 (1992)] and a phage from a clinical isolate [D. Lim, Mol. Microbiol. 6, 3531 (1992)] also integrate into thrW.
106. P22 [D. F. Lindsey, C. Martinez, J. R. Walker, J. Bacteriol. 174, 3834 (1992)] and a phage from a clinical isolate [D. Lim, Mol. Microbiol. 6, 3531 (1992)] also integrate into thrW.
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108. E. Kofoid and J. Roth, personal communication.
109. This is Laboratory of Genetics paper 3487. We thank the entire E. coli community for their support, encouragement, and sharing of data, and especially D. L. Daniels and N. Peterson, who were present at the creation. We also thank R. Straussburg and M. Guyer, our program administrators; R. R. Burgess and M. Sussman for critical reading of the manuscript; M. Borodovsky and W. S. Hayes for application of a new version of the GeneMark program to the analysis of the sequence; K. Rudd for his Ecoseq7 melds of GenBank data; J. Mahillon for providing I-Sce I strains; J. Roth and E. Kofoid for unpublished Salmonella data; the Japanese group under H. Mori and T. Horiuchi for cooperative competition; G. Pósfai and W. Szybalski for the popout strains; S. Baldwin, C. Allex, N. Manola, G. Bouriakov, and J. Schroeder of DNASTAR for extraordinary software; A. Huerta, H. Salgado, and D. Thieffry for help with promoter, operon, and regulatory site identification; T. Thiesen for Postscript illustrations; H. Kijenski, G. Peyrot, P. Soni, G. Diarra, E. Grotbeck, T. Forsythe, M. Maguire, M. Federle, S. Subramanian, and K. Kadner for excellent technical work; and 169 University of Wisconsin undergraduates who participated over the last decade. Supported by NIH grants P01 HG01428 (from the Human Genome Project) and S10 RR10379 (for ABI machines from the National Center for Research Resources-Biomedical Research Support Shared Instrumentation Grant). We thank IBM for the gift of workstations, the State of Wisconsin for remodeling support, and especially SmithKline Beecham Pharmaceutical and Genome Therapeutics Corp. for financial support of the annotation of this sequence. N.P. is an NSF fellow in molecular evolution.