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. 1992 May;58(5):1670–1676. doi: 10.1128/aem.58.5.1670-1676.1992

Conjugal transfer in Lactococcus lactis of a 68-kilobase-pair chromosomal fragment containing the structural gene for the peptide bacteriocin nisin.

T Gireesh 1, B E Davidson 1, A J Hillier 1
PMCID: PMC195656  PMID: 1622237

Abstract

Nisin-producing transconjugants were generated by mating nisin-producing strains of Lactococcus lactis subsp. lactis with derivatives of L. lactis subsp. lactis LM0230. The sucrose-utilizing ability and reduced bacteriophage sensitivity were also transferred with the nisin-producing character. Pulsed-field gel electrophoretic analysis of genomic DNA from donor, recipient, and nisin-producing transconjugants indicated that 68 kbp of DNA was transferred from the chromosome of the donor into the chromosome of the recipient in the conjugation process. The location of the transferred nisin structural gene spaN in the transconjugant HID500 was not stable, and cultures of strain HID500 were a mixture of different genotypes in which spaN was located at different positions in the chromosome on different SmaI fragments. ApaI, BglI, BssHII, NciI, SalI, and SmaI digests of genomic DNA were used to map the location of spaN in a donor (DL11) and a nisin-producing transconjugant (HID504).

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Selected References

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  1. Anderson D. G., McKay L. L. Simple and rapid method for isolating large plasmid DNA from lactic streptococci. Appl Environ Microbiol. 1983 Sep;46(3):549–552. doi: 10.1128/aem.46.3.549-552.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benkerroum N., Sandine W. E. Inhibitory action of nisin against Listeria monocytogenes. J Dairy Sci. 1988 Dec;71(12):3237–3245. doi: 10.3168/jds.S0022-0302(88)79929-4. [DOI] [PubMed] [Google Scholar]
  3. Broadbent J. R., Kondo J. K. Genetic construction of nisin-producing Lactococcus lactis subsp. cremoris and analysis of a rapid method for conjugation. Appl Environ Microbiol. 1991 Feb;57(2):517–524. doi: 10.1128/aem.57.2.517-524.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buchman G. W., Banerjee S., Hansen J. N. Structure, expression, and evolution of a gene encoding the precursor of nisin, a small protein antibiotic. J Biol Chem. 1988 Nov 5;263(31):16260–16266. [PubMed] [Google Scholar]
  5. Dodd H. M., Horn N., Gasson M. J. Analysis of the genetic determinant for production of the peptide antibiotic nisin. J Gen Microbiol. 1990 Mar;136(3):555–566. doi: 10.1099/00221287-136-3-555. [DOI] [PubMed] [Google Scholar]
  6. Donkersloot J. A., Thompson J. Simultaneous loss of N5-(carboxyethyl)ornithine synthase, nisin production, and sucrose-fermenting ability by Lactococcus lactis K1. J Bacteriol. 1990 Jul;172(7):4122–4126. doi: 10.1128/jb.172.7.4122-4126.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  8. Gonzalez C. F., Kunka B. S. Transfer of Sucrose-Fermenting Ability and Nisin Production Phenotype among Lactic Streptococci. Appl Environ Microbiol. 1985 Mar;49(3):627–633. doi: 10.1128/aem.49.3.627-633.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gross E., Kiltz H. H. The number and nature of , -unsaturated amino acids in subtilin. Biochem Biophys Res Commun. 1973 Jan 23;50(2):559–565. doi: 10.1016/0006-291x(73)90876-0. [DOI] [PubMed] [Google Scholar]
  10. Gross E. alpha, beta-Unsaturated and related amino acids in peptides and proteins. Adv Exp Med Biol. 1977;86B:131–153. doi: 10.1007/978-1-4757-9113-6_9. [DOI] [PubMed] [Google Scholar]
  11. Horn N., Swindell S., Dodd H., Gasson M. Nisin biosynthesis genes are encoded by a novel conjugative transposon. Mol Gen Genet. 1991 Aug;228(1-2):129–135. doi: 10.1007/BF00282457. [DOI] [PubMed] [Google Scholar]
  12. Kaletta C., Entian K. D. Nisin, a peptide antibiotic: cloning and sequencing of the nisA gene and posttranslational processing of its peptide product. J Bacteriol. 1989 Mar;171(3):1597–1601. doi: 10.1128/jb.171.3.1597-1601.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kellner R., Jung G., Hörner T., Zähner H., Schnell N., Entian K. D., Götz F. Gallidermin: a new lanthionine-containing polypeptide antibiotic. Eur J Biochem. 1988 Oct 15;177(1):53–59. doi: 10.1111/j.1432-1033.1988.tb14344.x. [DOI] [PubMed] [Google Scholar]
  14. LeBlanc D. J., Crow V. L., Lee L. N., Garon C. F. Influence of the lactose plasmid on the metabolism of galactose by Streptococcus lactis. J Bacteriol. 1979 Feb;137(2):878–884. doi: 10.1128/jb.137.2.878-884.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McKay L. L., Baldwin K. A., Zottola E. A. Loss of lactose metabolism in lactic streptococci. Appl Microbiol. 1972 Jun;23(6):1090–1096. doi: 10.1128/am.23.6.1090-1096.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Murphy M. C., Steele J. L., Daly C., McKay L. L. Concomitant conjugal transfer of reduced-bacteriophage-sensitivity mechanisms with lactose- and sucrose-fermenting ability in lactic streptococci. Appl Environ Microbiol. 1988 Aug;54(8):1951–1956. doi: 10.1128/aem.54.8.1951-1956.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Powell I. B., Ward A. C., Hillier A. J., Davidson B. E. Simultaneous conjugal transfer in Lactococcus to genes involved in bacteriocin production and reduced susceptibility to bacteriophages. FEMS Microbiol Lett. 1990 Oct;60(1-2):209–213. doi: 10.1016/0378-1097(90)90373-x. [DOI] [PubMed] [Google Scholar]
  18. Powell Ian B., Achen Marc G., Hillier Alan J., Davidson Barrie E. A Simple and Rapid Method for Genetic Transformation of Lactic Streptococci by Electroporation. Appl Environ Microbiol. 1988 Mar;54(3):655–660. doi: 10.1128/aem.54.3.655-660.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rauch P. J., Beerthuyzen M. M., de Vos W. M. Nucleotide sequence of IS904 from Lactococcus lactis subsp. lactis strain NIZO R5. Nucleic Acids Res. 1990 Jul 25;18(14):4253–4254. doi: 10.1093/nar/18.14.4253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reed K. C., Mann D. A. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. doi: 10.1093/nar/13.20.7207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schnell N., Entian K. D., Schneider U., Götz F., Zähner H., Kellner R., Jung G. Prepeptide sequence of epidermin, a ribosomally synthesized antibiotic with four sulphide-rings. Nature. 1988 May 19;333(6170):276–278. doi: 10.1038/333276a0. [DOI] [PubMed] [Google Scholar]
  22. Steele J. L., McKay L. L. Partial characterization of the genetic basis for sucrose metabolism and nisin production in Streptococcus lactis. Appl Environ Microbiol. 1986 Jan;51(1):57–64. doi: 10.1128/aem.51.1.57-64.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Steen M. T., Chung Y. J., Hansen J. N. Characterization of the nisin gene as part of a polycistronic operon in the chromosome of Lactococcus lactis ATCC 11454. Appl Environ Microbiol. 1991 Apr;57(4):1181–1188. doi: 10.1128/aem.57.4.1181-1188.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tanskanen E. I., Tulloch D. L., Hillier A. J., Davidson B. E. Pulsed-Field Gel Electrophoresis of SmaI Digests of Lactococcal Genomic DNA, a Novel Method of Strain Identification. Appl Environ Microbiol. 1990 Oct;56(10):3105–3111. doi: 10.1128/aem.56.10.3105-3111.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Terzaghi B. E., Sandine W. E. Improved medium for lactic streptococci and their bacteriophages. Appl Microbiol. 1975 Jun;29(6):807–813. doi: 10.1128/am.29.6.807-813.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Thompson J., Nguyen N. Y., Sackett D. L., Donkersloot J. A. Transposon-encoded sucrose metabolism in Lactococcus lactis. Purification of sucrose-6-phosphate hydrolase and genetic linkage to N5-(L-1-carboxyethyl)-L-ornithine synthase in strain K1. J Biol Chem. 1991 Aug 5;266(22):14573–14579. [PubMed] [Google Scholar]
  27. Tsai H. J., Sandine W. E. Conjugal transfer of nisin plasmid genes from Streptococcus lactis 7962 to Leuconostoc dextranicum 181. Appl Environ Microbiol. 1987 Feb;53(2):352–357. doi: 10.1128/aem.53.2.352-357.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tulloch D. L., Finch L. R., Hillier A. J., Davidson B. E. Physical map of the chromosome of Lactococcus lactis subsp. lactis DL11 and localization of six putative rRNA operons. J Bacteriol. 1991 May;173(9):2768–2775. doi: 10.1128/jb.173.9.2768-2775.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vollrath D., Davis R. W. Resolution of DNA molecules greater than 5 megabases by contour-clamped homogeneous electric fields. Nucleic Acids Res. 1987 Oct 12;15(19):7865–7876. doi: 10.1093/nar/15.19.7865. [DOI] [PMC free article] [PubMed] [Google Scholar]

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