Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1987 Jul;53(7):1584–1588. doi: 10.1128/aem.53.7.1584-1588.1987

Cloning of a Streptococcus lactis subsp. lactis Chromosomal Fragment Associated with the Ability To Grow in Milk

Atte von Wright 1,*, Soile Tynkkynen 1, Martin Suominen 1
PMCID: PMC203914  PMID: 16347386

Abstract

A chromosomal fragment of 6.7 megadaltons (MDa), apparently containing the genes for milk protein utilization by Streptococcus lactis subsp. lactis SSL135, was cloned in S. lactis subsp. lactis MG1614, a proteinase-negative strain. For the cloning, the chromosomal DNA of SSL135 was cleaved with restriction enzyme BamHI and the resulting fragments were ligated to the single BclI site of pVS2, a 3.3-MDa chloramphenicol-erythromycin double-resistance plasmid constructed in this laboratory. S. lactis subsp. lactis MG1614 was transformed by using this ligation mixture and selecting for chloramphenicol resistance and growth in citrated milk medium. One clone containing a 10.0-MDa plasmid, subsequently designated as pVS6, was chosen for further studies. Despite the lack of homology with previously characterized proteinase genes of lactic streptococci, the cloned insert consistently conveyed the ability to grow in milk to proteinase-negative recipients in repeated transformation experiments. The genetic evidence suggests that the main part of the gene(s) for the proposed proteinase activity is located within a 3.8-MDa BglII fragment of the clone.

Full text

PDF
1584

Images in this article

1587Image
on p.1587

Selected References

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

  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. Behnke D., Gilmore M. S. Location of antibiotic resistance determinants, copy control, and replication functions on the double-selective streptococcal cloning vector pGB301. Mol Gen Genet. 1981;184(1):115–120. doi: 10.1007/BF00271206. [DOI] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Gasson M. J. Plasmid complements of Streptococcus lactis NCDO 712 and other lactic streptococci after protoplast-induced curing. J Bacteriol. 1983 Apr;154(1):1–9. doi: 10.1128/jb.154.1.1-9.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibodies. J Bacteriol. 1982 May;150(2):804–814. doi: 10.1128/jb.150.2.804-814.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kok J., van Dijl J. M., van der Vossen J. M., Venema G. Cloning and expression of a Streptococcus cremoris proteinase in Bacillus subtilis and Streptococcus lactis. Appl Environ Microbiol. 1985 Jul;50(1):94–101. doi: 10.1128/aem.50.1.94-101.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kok J., van der Vossen J. M., Venema G. Construction of plasmid cloning vectors for lactic streptococci which also replicate in Bacillus subtilis and Escherichia coli. Appl Environ Microbiol. 1984 Oct;48(4):726–731. doi: 10.1128/aem.48.4.726-731.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Law B. A., Kolstad J. Proteolytic systems in lactic acid bacteria. Antonie Van Leeuwenhoek. 1983 Sep;49(3):225–245. doi: 10.1007/BF00399500. [DOI] [PubMed] [Google Scholar]
  9. Marinus M. G. Location of DNA methylation genes on the Escherichia coli K-12 genetic map. Mol Gen Genet. 1973 Dec 14;127(1):47–55. doi: 10.1007/BF00267782. [DOI] [PubMed] [Google Scholar]
  10. McKay L. L. Functional properties of plasmids in lactic streptococci. Antonie Van Leeuwenhoek. 1983 Sep;49(3):259–274. doi: 10.1007/BF00399502. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. von Wright A., Taimisto A. M., Sivelä S. Effect of Ca2+ ions on plasmid transformation of Streptococcus lactis protoplasts. Appl Environ Microbiol. 1985 Oct;50(4):1100–1102. doi: 10.1128/aem.50.4.1100-1102.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES