Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 Sep;175(17):5420–5427. doi: 10.1128/jb.175.17.5420-5427.1993

Genetic analysis of microcin H47 antibiotic system.

C Gaggero 1, F Moreno 1, M Laviña 1
PMCID: PMC206597  PMID: 8366029

Abstract

The microcin H47 genetic determinants span a DNA region of ca. 10 kb and represent the first description of an enterobacterial antibiotic system located in the chromosome of the producing strain. Transcriptional and translational fusions to lacZ showed a complex transcriptional organization of the microcin H47 system. Complementation tests identified six genes that are necessary for the production of the antibiotic; the products of two of them are involved in the export of microcin to the extracellular medium. The immunity determinant was located in an 0.8-kb DNA fragment. There is a putative "silent region" of ca. 3 kb inside the system that could not be clearly related to any antibiotic function. Protein products were identified and assigned to three production genes and also to a gene from the silent region.

Full text

PDF
5420

Images in this article

5425Image
on p.5425

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  3. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Garrido M. C., Herrero M., Kolter R., Moreno F. The export of the DNA replication inhibitor Microcin B17 provides immunity for the host cell. EMBO J. 1988 Jun;7(6):1853–1862. doi: 10.1002/j.1460-2075.1988.tb03018.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Genilloud O., Moreno F., Kolter R. DNA sequence, products, and transcriptional pattern of the genes involved in production of the DNA replication inhibitor microcin B17. J Bacteriol. 1989 Feb;171(2):1126–1135. doi: 10.1128/jb.171.2.1126-1135.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gilson L., Mahanty H. K., Kolter R. Four plasmid genes are required for colicin V synthesis, export, and immunity. J Bacteriol. 1987 Jun;169(6):2466–2470. doi: 10.1128/jb.169.6.2466-2470.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gilson L., Mahanty H. K., Kolter R. Genetic analysis of an MDR-like export system: the secretion of colicin V. EMBO J. 1990 Dec;9(12):3875–3884. doi: 10.1002/j.1460-2075.1990.tb07606.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hiraga S., Niki H., Ogura T., Ichinose C., Mori H., Ezaki B., Jaffé A. Chromosome partitioning in Escherichia coli: novel mutants producing anucleate cells. J Bacteriol. 1989 Mar;171(3):1496–1505. doi: 10.1128/jb.171.3.1496-1505.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  10. Kolter R., Moreno F. Genetics of ribosomally synthesized peptide antibiotics. Annu Rev Microbiol. 1992;46:141–163. doi: 10.1146/annurev.mi.46.100192.001041. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Laviña M., Gaggero C., Moreno F. Microcin H47, a chromosome-encoded microcin antibiotic of Escherichia coli. J Bacteriol. 1990 Nov;172(11):6585–6588. doi: 10.1128/jb.172.11.6585-6588.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leach D., Symonds N. The isolation and characterisation of a plaque-forming derivative of bacteriophage Mu carrying a fragment of Tn3 conferring ampicillin resistance. Mol Gen Genet. 1979 May 4;172(2):179–184. doi: 10.1007/BF00268280. [DOI] [PubMed] [Google Scholar]
  14. Manoil C. Analysis of protein localization by use of gene fusions with complementary properties. J Bacteriol. 1990 Feb;172(2):1035–1042. doi: 10.1128/jb.172.2.1035-1042.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  16. Novoa M. A., Díaz-Guerra L., San Millán J. L., Moreno F. Cloning and mapping of the genetic determinants for microcin C7 production and immunity. J Bacteriol. 1986 Dec;168(3):1384–1391. doi: 10.1128/jb.168.3.1384-1391.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wandersman C., Delepelaire P. TolC, an Escherichia coli outer membrane protein required for hemolysin secretion. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4776–4780. doi: 10.1073/pnas.87.12.4776. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES