1887

Abstract

K-12 was originally isolated 100 years ago and since then it has become an invaluable model organism and a cornerstone of molecular biology research. However, despite its pedigree, since its initial isolation K-12 has been repeatedly cultured, passaged and mutagenized, resulting in an organism that carries many genetic changes. To understand more about this important model organism, we have sequenced the genomes of two ancestral K-12 strains, WG1 and EMG2, considered to be the progenitors of many key laboratory strains. Our analysis confirms that these strains still carry genetic elements such as bacteriophage lambda (λ) and the F plasmid, but also indicates that they have undergone extensive laboratory-based evolution. Thus, scrutinizing the genomes of ancestral K-12 strains leads us to examine whether K-12 is a sufficiently robust model organism for 21st century microbiology.

Funding
This study was supported by the:
  • Biotechnology and Biological Sciences Research Council (Award BB/E01044X/1)
    • Principle Award Recipient: JonL Hobman
  • Biotechnology and Biological Sciences Research Council (Award BB/R017689/1 and BB/W00285X/1)
    • Principle Award Recipient: StephenJW Busby
  • Biotechnology and Biological Sciences Research Council (Award BB/R017689/1 and BB/W00285X/1)
    • Principle Award Recipient: DouglasF Browning
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/mgen/10.1099/mgen.0.000922
2023-02-06
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/mgen/9/2/mgen000922.html?itemId=/content/journal/mgen/10.1099/mgen.0.000922&mimeType=html&fmt=ahah

References

  1. Bachmann BJ. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 1972; 36:525–557 [View Article] [PubMed]
    [Google Scholar]
  2. Clowes RC, Hayes W. Experiments in Microbial Genetics Oxford, Edinburgh: Blackwell Scientific Publications; 1968
    [Google Scholar]
  3. Hobman JL, Penn CW, Pallen MJ. Laboratory strains of Escherichia coli: model citizens or deceitful delinquents growing old disgracefully?. Mol Microbiol 2007; 64:881–885 [View Article]
    [Google Scholar]
  4. Blattner FR, Plunkett G, Bloch CA, Perna NT, Burland V et al. The complete genome sequence of Escherichia coli K-12. Science 1997; 277:1453–1462 [View Article]
    [Google Scholar]
  5. Carattoli A, Zankari E, García-Fernández A, Voldby Larsen M, Lund O et al. In silico detection and typing of plasmids using PlasmidFinder and plasmid multilocus sequence typing. Antimicrob Agents Chemother 2014; 58:3895–3903 [View Article] [PubMed]
    [Google Scholar]
  6. Casjens SR, Hendrix RW. Bacteriophage lambda: early pioneer and still relevant. Virology 2015; 479–480:310–330 [View Article]
    [Google Scholar]
  7. Koraimann G. Spread and persistence of virulence and antibiotic resistance genes: a ride on the F plasmid conjugation module. EcoSal Plus 2018; 8:ESP-0003-2018 [View Article]
    [Google Scholar]
  8. Liu D, Reeves PR. Escherichia coli K12 regains its O antigen. Microbiology 1994; 140:49–57 [View Article]
    [Google Scholar]
  9. Stevenson G, Neal B, Liu D, Hobbs M, Packer NH et al. Structure of the O antigen of Escherichia coli K-12 and the sequence of its rfb gene cluster. J Bacteriol 1994; 176:4144–4156 [View Article] [PubMed]
    [Google Scholar]
  10. Smith HW. Survival of orally administered E. coli K 12 in alimentary tract of man. Nature 1975; 255:500–502 [View Article] [PubMed]
    [Google Scholar]
  11. Anderson ES. Viability of, and transfer of a plasmid from, E. coli K12 in human intestine. Nature 1975; 255:502–504 [View Article] [PubMed]
    [Google Scholar]
  12. Hayashi K, Morooka N, Yamamoto Y, Fujita K, Isono K et al. Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110. Mol Syst Biol 2006; 2:2006.0007 [View Article]
    [Google Scholar]
  13. Dunne KA, Chaudhuri RR, Rossiter AE, Beriotto I, Browning DF et al. Sequencing a piece of history: complete genome sequence of the original Escherichia coli strain. Microb Genom 2017; 3:000106 [View Article]
    [Google Scholar]
  14. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for illumina sequence data. Bioinformatics 2014; 30:2114–2120 [View Article] [PubMed]
    [Google Scholar]
  15. Wick RR, Judd LM, Gorrie CL, Holt KE. Unicycler: resolving bacterial genome assemblies from short and long sequencing reads. PLoS Comput Biol 2017; 13:e1005595 [View Article]
    [Google Scholar]
  16. Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics 2014; 30:2068–2069 [View Article]
    [Google Scholar]
  17. Rutherford K, Parkhill J, Crook J, Horsnell T, Rice P et al. Artemis: sequence visualization and annotation. Bioinformatics 2000; 16:944–945 [View Article]
    [Google Scholar]
  18. Carver TJ, Rutherford KM, Berriman M, Rajandream M-A, Barrell BG et al. ACT: the Artemis Comparison Tool. Bioinformatics 2005; 21:3422–3423 [View Article]
    [Google Scholar]
  19. Grant JR, Stothard P. The CGView server: a comparative genomics tool for circular genomes. Nucleic Acids Res 2008; 36:W181–W184 [View Article]
    [Google Scholar]
  20. Zhang Z, Schwartz S, Wagner L, Miller W. A greedy algorithm for aligning DNA sequences. J Comput Biol 2000; 7:203–214 [View Article]
    [Google Scholar]
  21. Hill CW, Harnish BW. Inversions between ribosomal RNA genes of Escherichia coli. Proc Natl Acad Sci USA 1981; 78:7069–7072 [View Article]
    [Google Scholar]
  22. Nobelmann B, Lengeler JW. Molecular analysis of the gat genes from Escherichia coli and of their roles in galactitol transport and metabolism. J Bacteriol 1996; 178:6790–6795 [View Article] [PubMed]
    [Google Scholar]
  23. Zhang Z, Kukita C, Humayun MZ, Saier MH. Environment-directed activation of the Escherichia coli flhDC operon by transposons. Microbiology 2017; 163:554–569 [View Article]
    [Google Scholar]
  24. Barker CS, Prüss BM, Matsumura P. Increased motility of Escherichia coli by insertion sequence element integration into the regulatory region of the flhD operon. J Bacteriol 2004; 186:7529–7537 [View Article]
    [Google Scholar]
  25. Naas T, Blot M, Fitch WM, Arber W. Insertion sequence-related genetic variation in resting Escherichia coli K-12. Genetics 1994; 136:721–730 [View Article] [PubMed]
    [Google Scholar]
  26. Naas T, Blot M, Fitch WM, Arber W. Dynamics of IS-related genetic rearrangements in resting Escherichia coli K-12. Mol Biol Evol 1995; 12:198–207 [View Article] [PubMed]
    [Google Scholar]
  27. Freddolino PL, Amini S, Tavazoie S. Newly identified genetic variations in common Escherichia coli MG1655 stock cultures. J Bacteriol 2012; 194:303–306 [View Article] [PubMed]
    [Google Scholar]
  28. Kuhnert P, Nicolet J, Frey J. Rapid and accurate identification of Escherichia coli K-12 strains. Appl Environ Microbiol 1995; 61:4135–4139 [View Article] [PubMed]
    [Google Scholar]
  29. Jeong H, Barbe V, Lee CH, Vallenet D, Yu DS et al. Genome sequences of Escherichia coli B strains REL606 and BL21(DE3). J Mol Biol 2009; 394:644–652 [View Article] [PubMed]
    [Google Scholar]
  30. Browning DF, Wells TJ, França FLS, Morris FC, Sevastsyanovich YR et al. Laboratory adapted Escherichia coli K-12 becomes a pathogen of Caenorhabditis elegans upon restoration of O antigen biosynthesis. Mol Microbiol 2013; 87:939–950 [View Article] [PubMed]
    [Google Scholar]
  31. Yen MR, Peabody CR, Partovi SM, Zhai Y, Tseng YH et al. Protein-translocating outer membrane porins of Gram-negative bacteria. Biochim Biophys Acta 2002; 1562:6–31 [View Article] [PubMed]
    [Google Scholar]
  32. Cheah KC, Skurray R. The F plasmid carries an IS3 insertion within finO. J Gen Microbiol 1986; 132:3269–3275 [View Article]
    [Google Scholar]
  33. Yoshioka Y, Ohtsubo H, Ohtsubo E. Repressor gene finO in plasmids R100 and F: constitutive transfer of plasmid F is caused by insertion of IS3 into F finO. J Bacteriol 1987; 169:619–623 [View Article]
    [Google Scholar]
  34. Sanger F, Coulson AR, Hong GF, Hill DF, Petersen GB. Nucleotide sequence of bacteriophage lambda DNA. J Mol Biol 1982; 162:729–773 [View Article] [PubMed]
    [Google Scholar]
  35. Hendrix RW, Duda RL. Bacteriophage lambda PaPa: not the mother of all lambda phages. Science 1992; 258:1145–1148 [View Article] [PubMed]
    [Google Scholar]
  36. Siegele DA, Hu JC, Gross CA. Mutations in rpoD, the gene encoding the sigma 70 subunit of Escherichia coli RNA polymerase, that increase expression of the lac operon in the absence of CAP-cAMP. J Mol Biol 1988; 203:29–37 [View Article] [PubMed]
    [Google Scholar]
  37. Siegele DA, Hu JC, Walter WA, Gross CA. Altered promoter recognition by mutant forms of the sigma 70 subunit of Escherichia coli RNA polymerase. J Mol Biol 1989; 206:591–603 [View Article] [PubMed]
    [Google Scholar]
  38. Caslake LF, Ashraf SI, Summers AO. Mutations in the alpha and sigma-70 subunits of RNA polymerase affect expression of the mer operon. J Bacteriol 1997; 179:1787–1795 [View Article] [PubMed]
    [Google Scholar]
  39. Jishage M, Kvint K, Shingler V, Nyström T. Regulation of sigma factor competition by the alarmone ppGpp. Genes Dev 2002; 16:1260–1270 [View Article] [PubMed]
    [Google Scholar]
  40. Cicero MP, Sharp MM, Gross CA, Kreuzer KN. Substitutions in bacteriophage T4 AsiA and Escherichia coli sigma(70) that suppress T4 motA activation mutations. J Bacteriol 2001; 183:2289–2297 [View Article] [PubMed]
    [Google Scholar]
  41. Lombardo MJ, Bagga D, Miller CG. Mutations in rpoA affect expression of anaerobically regulated genes in Salmonella typhimurium. J Bacteriol 1991; 173:7511–7518 [View Article] [PubMed]
    [Google Scholar]
  42. Schaub RE, Hayes CS. Deletion of the RluD pseudouridine synthase promotes SsrA peptide tagging of ribosomal protein S7. Mol Microbiol 2011; 79:331–341 [View Article] [PubMed]
    [Google Scholar]
  43. Pavlov MY, Freistroffer DV, Dincbas V, MacDougall J, Buckingham RH et al. A direct estimation of the context effect on the efficiency of termination. J Mol Biol 1998; 284:579–590 [View Article] [PubMed]
    [Google Scholar]
  44. Collier J, Binet E, Bouloc P. Competition between SsrA tagging and translational termination at weak stop codons in Escherichia coli. Mol Microbiol 2002; 45:745–754 [View Article] [PubMed]
    [Google Scholar]
  45. Dinçbas-Renqvist V, Engström A, Mora L, Heurgué-Hamard V, Buckingham R et al. A post-translational modification in the GGQ motif of RF2 from Escherichia coli stimulates termination of translation. EMBO J 2000; 19:6900–6907 [View Article] [PubMed]
    [Google Scholar]
  46. Uno M, Ito K, Nakamura Y. Functional specificity of amino acid at position 246 in the tRNA mimicry domain of bacterial release factor 2. Biochimie 1996; 78:935–943 [View Article] [PubMed]
    [Google Scholar]
  47. Jensen KF. The Escherichia coli K-12 “wild types” W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels. J Bacteriol 1993; 175:3401–3407 [View Article] [PubMed]
    [Google Scholar]
  48. Soupene E, van Heeswijk WC, Plumbridge J, Stewart V, Bertenthal D et al. Physiological studies of Escherichia coli strain MG1655: growth defects and apparent cross-regulation of gene expression. J Bacteriol 2003; 185:5611–5626 [View Article] [PubMed]
    [Google Scholar]
  49. Lawther RP, Calhoun DH, Gray J, Adams CW, Hauser CA et al. DNA sequence fine-structure analysis of ilvG (IlvG+) mutations of Escherichia coli K-12. J Bacteriol 1982; 149:294–298 [View Article] [PubMed]
    [Google Scholar]
  50. Schuster S, Vavra M, Greim L, Kern WV. Exploring the contribution of the AcrB homolog MdtF to drug resistance and dye efflux in a multidrug resistant E. coli isolate. Antibiotics 2021; 10:503 [View Article]
    [Google Scholar]
  51. Weiss B. Endonuclease V of Escherichia coli prevents mutations from nitrosative deamination during nitrate/nitrite respiration. Mutat Res 2001; 461:301–309 [View Article] [PubMed]
    [Google Scholar]
  52. Zimmer C. Microcosm: E. coli and the New Science of Life New York: Pantheon Books; 2008
    [Google Scholar]
  53. Neidhardt FC, Curtiss R. Escherichia coli and Salmonella: Cellular and Molecular Biology Washington, DC: American Society for Microbiology; 1996
    [Google Scholar]
  54. Brown SD, Jun S. Complete genome sequence of Escherichia coli NCM3722. Genome Announc 2015; 3:e00879-15 [View Article]
    [Google Scholar]
  55. Rand JM, Gordon GC, Mehrer CR, Pfleger BF. Genome sequence and analysis of Escherichia coli production strain LS5218. Metab Eng Commun 2017; 5:78–83 [View Article] [PubMed]
    [Google Scholar]
  56. Nahku R, Peebo K, Valgepea K, Barrick JE, Adamberg K et al. Stock culture heterogeneity rather than new mutational variation complicates short-term cell physiology studies of Escherichia coli K-12 MG1655 in continuous culture. Microbiology 2011; 157:2604–2610 [View Article]
    [Google Scholar]
  57. Spira B, de Almeida Toledo R, Maharjan RP, Ferenci T. The uncertain consequences of transferring bacterial strains between laboratories – rpoS instability as an example. BMC Microbiol 2011; 11:248 [View Article]
    [Google Scholar]
  58. Hooton SPT, Pritchard ACW, Asiani K, Gray-Hammerton CJ, Stekel DJ et al. Laboratory stock variants of the archetype silver resistance plasmid pMG101 demonstrate plasmid fusion, loss of transmissibility, and transposition of Tn7/pco/sil into the host chromosome. Front Microbiol 2021; 12:723322 [View Article]
    [Google Scholar]
  59. Jishage M, Ishihama A. Variation in RNA polymerase sigma subunit composition within different stocks of Escherichia coli W3110. J Bacteriol 1997; 179:959–963 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/mgen/10.1099/mgen.0.000922
Loading
/content/journal/mgen/10.1099/mgen.0.000922
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Supplementary material 2

EXCEL
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error