1887

Abstract

We have identified mutations in three different chromosomal genes of K12 which reduce sensitivity to microcin B17. Mutations in and genes affected production of an outer membrane porin protein, OmpF, and resulted in reduced sensitivity to a number of other agents (colicins, bacteriophages) besides microcin B17. The third class of mutants were specifically and highly resistant to microcin B17. The mutations in these strains were mapped to a gene (), located at 8·7 min on the K12 chromosome, which is closely linked to The wild-type allele was cloned into multiple copy number plasmids, and its location within the cloned DNA fragment was further defined by mutagenesis with MiniMudII1681. These insertion mutations resulted in in-frame fusions between the and genes, thereby allowing us to determine the direction of gene transcription. Plasmids carrying these gene fusions produced low levels of β-galactosidase, indicating that the gene is poorly expressed. We have been unable to identify the gene product, but indirect evidence indicates that it might be an envelope protein involved in microcin uptake.

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1986-06-01
2021-08-05
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References

  1. Asensio C., Perez Diaz J. C., Martinez M. C., Baquero F. 1976; A new family of low molecular weight antibiotics from Enterobacteria.. Biochemical and Biophysical Research Communications 69:7–14
    [Google Scholar]
  2. Bachmann B. J. 1983; Linkage map of Escherichia coli K12 Edition 7. Microbiological Reviews 47:180–230
    [Google Scholar]
  3. Baquero F., Moreno F. 1984; The microcins.. FEMS Microbiology Letters 23:117–124
    [Google Scholar]
  4. Baquero F., Bouanchaud D., Martinez M. C., Fernandez C. 1978; Microcin plasmids: a group of extrachromosomal elements coding for low- molecular-weight antibiotics in Escherichia coli.. Journal of Bacteriology 135:342–347
    [Google Scholar]
  5. Berg D. E. 1977; Insertion and excision of a transposable kanamycin resistance determinant Tn 5DNA . in Insertion Elements, Plasmids and Episomes pp. 205–212 Editor Bukhari A.I, Shapiro J. A., Adhya. S. L. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  6. Berg P. E. 1981; Cloning and characterization of the Escherichia coli gene coding for the alkaline phosphatase.. Journal of Bacteriology 146:660–667
    [Google Scholar]
  7. Boidol W., Simonis M., Topert M., Siewert G. 1982; Recombinant plasmids with genes for the biosynthesis of alkaline phosphatase of Escherichia coli.. Molecular and General Genetics 185:510–512
    [Google Scholar]
  8. Calos M. P., Miller J. H. 1981; The DNA sequence change resulting from the lacl4' mutation which greatly increases promoter strength.. Molecular and General Genetics 183:559–560
    [Google Scholar]
  9. Casadaban M. 1976; Transposition and fusion of the lac genes to selected promoters in E. coli using bacteriophage lambda and Mu.. Journal of Molecular Biology 104:541–555
    [Google Scholar]
  10. Castilho B. A., Olfson P., Casadaban M. J. 1984; Plasmid insertion mutagenesis and lac gene fusion with mini-Mu bacteriophage transposons.. Journal of Bacteriology 158:488–495
    [Google Scholar]
  11. Clarke L., Carbon J. 1976; A colony bank containing synthetic ColEl hybrid plasmids representative of the entire Escherichia coli genome.. Cell 9:91–99
    [Google Scholar]
  12. DATTA D. B., Arden B., Henning U. 1977; Major proteins of the Escherichia coli outer cell envelope membrane as bacteriophage receptors.. Journal of Bacteriology 131:821–829
    [Google Scholar]
  13. DAVIS R. W., BOTSTEIN D., ROTH J. R. 1980; Advanced Bacterial Genetics.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  14. Hall M. N., Silhavy T. J. 1979; Transcriptional regulation of Escherichia coli K12 major outer membrane protein lb.. Journal of Bacteriology 140:342–350
    [Google Scholar]
  15. Hall M. N., Silhavy T. J. 1981; The ompB locus and the regulation of the major outer membrane porin proteins of Escherichia coli K 12 . Journal of Molecular Biology 146:23–43
    [Google Scholar]
  16. HANKTE K. 1978; Major outer membrane proteins of Escherichia coli K12 serve as receptors for the phage T2 (protein la) and 434 (protein lb).. Molecular and General Genetics 164:131–135
    [Google Scholar]
  17. Hernandez-Chico C., Herrero M., Rejas M., Millan J. L., San Moreno F. 1982; Gene ompR and regulation of microcin 17 and colicin E2 syntheses.. Journal of Bacteriology 152:897–900
    [Google Scholar]
  18. HERRERO M. 1984; Estudios sobre elmodo de accion de la microcina B17 . PhD thesis, Universidad Complu- tense de Madrid, Spain.
    [Google Scholar]
  19. Herrero M., Moreno F. 1986; Microcin B17 blocks DNA replication and induces the SOS system in Escherichia coli.. Journal of General Microbiology 132:393–402
    [Google Scholar]
  20. Herrero M., Kolter R., Moreno F. 1986; Effects of microcin B17 on microcin B17 immune cells. Journal of General Microbiology 132:403–410
    [Google Scholar]
  21. Inouye H., Michaelis S., Wright A., Beckwith J. 1981; Cloning and restriction mapping of the alkaline phosphatase structural gene (phoA) of Escherichia coli and generation of deletion mutants “in vitro”.. Journal of Bacteriology 146:668–675
    [Google Scholar]
  22. Jorgensen R. A., Rothstein S. J., Reznikoff W. S. 1979; A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance.. Molecular and General Genetics 177:65–72
    [Google Scholar]
  23. Konisky J. 1979; Specific transport systems and receptors for colicins and phages.. in Bacterial Outer Membranes. Biogenesis and Functions. Editor Inouye. M. New York: Wiley.;
    [Google Scholar]
  24. Low K. B. 1972; Escherichia coliK12 F-prime factors old and new. . Bacteriological Reviews 36:587–607
    [Google Scholar]
  25. Maniatis T., Fritsch E. F., Sambrook J. 1982; Molecular Cloning: a Laboratory Manual.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  26. Mayr-Harting A., Hedges A. J., Berkeley R. C. W. 1972; Methods for studying bacteriocins.. Methods in Microbiology 7A:315–422
    [Google Scholar]
  27. Messing J. 1983; New M13 vectors for cloning. Methods in Enzvmology 101:20–78
    [Google Scholar]
  28. Miller J. H. 1972; Experiments in Molecular Genetics.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  29. Mock M., Pugsley A. P. 1982; The BtuB group Col plasmids and homology between the colicins they encode.. Journal of Bacteriology 150:1069–1076
    [Google Scholar]
  30. Neidhardt F. C., Bloch P. L., Smith D. F. 1974; Culture medium for Enterobacteria.. Journal of Bacteriology 119:736–747
    [Google Scholar]
  31. O'Conner H. B., Malamy M. H. 1983; A new insertion sequence, IS 121 is found on the Mud II (Ap lac) bacteriophage and the Escherichia coli chromosome.. Journal of Bacteriology 156:669–679
    [Google Scholar]
  32. Pugsley A. P. 1983; Autoinduced synthesis of colicin E2. Molecular and General Genetics 190:379–383
    [Google Scholar]
  33. Pugsley A. P. 1984; The ins and outs of colicins. Parts I and II.. Microbiological Sciences 1:168–205
    [Google Scholar]
  34. PUGSLEY A. P. 1985; Escherichia coli K12 strains for use in the identification and characterization of colicins.. Journal of General Microbiology 131:369–376
    [Google Scholar]
  35. Pugsley A. P., Schwartz M., Lavina M., Moreno F. 1983; On the effect of ompR on colicin E2 production. . FEMS Microbiology Letters 19:87–92
    [Google Scholar]
  36. Sanger F., Coulson A. R., Friedmann T., Air G. M., Barrell B. G., Brown N. L., Fidoles J. C., Autchison iii C. A., Slocombe P. H., Smith M. 1978; The nucleotide sequence of bacteriophage (f)X 174. Journal of Molecular Biology 125:225–246
    [Google Scholar]
  37. Sanger F., Coulson A. R., Hong G. F., Hill D. F., Petersen G. B. 1982; Nucleotide sequence of bacteriophage lambda DNA.. Journal of Molecular Biology 162:729–773
    [Google Scholar]
  38. San Millan J. L., Hernandez-Chico C., Pereda P., Moreno F. 1985a; Cloning and mapping of the genetic determinants for microcin B17 production and immunity.. Journal of Bacteriology 163:275–281
    [Google Scholar]
  39. San Millan J. L., Kolter R., Moreno F. 1985; Plasmid genes required for microcin B17 production.. Journal of Bacteriology 163:1016–1020
    [Google Scholar]
  40. Sarthy A., Michaelis S., Beckwith J. 1981; Use of gene fusions to determine the orientation of the phoA gene on the Escherichia coli chromosome.. Journal of Bacteriology 145:293–298
    [Google Scholar]
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