Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1991 Feb;57(2):333–340. doi: 10.1128/aem.57.2.333-340.1991

Isolation and characterization of Lactococcus lactis subsp. lactis promoters.

T Koivula 1, M Sibakov 1, I Palva 1
PMCID: PMC182715  PMID: 1707605

Abstract

DNA fragments with promoter activity were isolated from the chromosome of Lactococcus lactis subsp. lactis. For the isolation, a promoter probe vector based on the cat gene was constructed, which allowed direct selection with chloramphenicol in Bacillus subtilis and L. lactis. Four of the putative promoters (P1, P2, P10, and P21) were analyzed further by sequencing, mapping of the 5' end of the mRNA, Northern (RNA blot) hybridization, and chloramphenicol acetyltransferase activity measurements. From these fragments, -10 and -35 regions resembling the consensus Escherichia coli sigma 70 and B. subtilis sigma 43 promoters were identified. Another set of promoters, together with a signal sequence, were also isolated from the same organism. These fragments promoted secretion of TEM beta-lactamase from L. lactis. When the two sets of promoters were compared, it was found that the ones isolated with the cat vector were more efficient (produced more mRNA). By changing the promoter part of the promoter-signal sequence fragment giving the best TEM beta-lactamase secretion into a more efficient one (P2), a 10-fold increase in enzyme production was obtained.

Full text

PDF
333

Images in this article

337Image
on p.337
337Image
on p.337

Selected References

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

  1. 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]
  2. Chopin M. C., Chopin A., Rouault A., Galleron N. Insertion and amplification of foreign genes in the Lactococcus lactis subsp. lactis chromosome. Appl Environ Microbiol. 1989 Jul;55(7):1769–1774. doi: 10.1128/aem.55.7.1769-1774.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Doi R. H., Wang L. F. Multiple procaryotic ribonucleic acid polymerase sigma factors. Microbiol Rev. 1986 Sep;50(3):227–243. doi: 10.1128/mr.50.3.227-243.1986. [DOI] [PMC free article] [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. Graves M. C., Rabinowitz J. C. In vivo and in vitro transcription of the Clostridium pasteurianum ferredoxin gene. Evidence for "extended" promoter elements in gram-positive organisms. J Biol Chem. 1986 Aug 25;261(24):11409–11415. [PubMed] [Google Scholar]
  6. Gryczan T. J., Contente S., Dubnau D. Characterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis. J Bacteriol. 1978 Apr;134(1):318–329. doi: 10.1128/jb.134.1.318-329.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  8. Hattori M., Sakaki Y. Dideoxy sequencing method using denatured plasmid templates. Anal Biochem. 1986 Feb 1;152(2):232–238. doi: 10.1016/0003-2697(86)90403-3. [DOI] [PubMed] [Google Scholar]
  9. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Holo H., Nes I. F. High-Frequency Transformation, by Electroporation, of Lactococcus lactis subsp. cremoris Grown with Glycine in Osmotically Stabilized Media. Appl Environ Microbiol. 1989 Dec;55(12):3119–3123. doi: 10.1128/aem.55.12.3119-3123.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Inamine J. M., Lee L. N., LeBlanc D. J. Molecular and genetic characterization of lactose-metabolic genes of Streptococcus cremoris. J Bacteriol. 1986 Sep;167(3):855–862. doi: 10.1128/jb.167.3.855-862.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kiwaki M., Ikemura H., Shimizu-Kadota M., Hirashima A. Molecular characterization of a cell wall-associated proteinase gene from Streptococcus lactis NCDO763. Mol Microbiol. 1989 Mar;3(3):359–369. doi: 10.1111/j.1365-2958.1989.tb00181.x. [DOI] [PubMed] [Google Scholar]
  13. Kok J., Leenhouts K. J., Haandrikman A. J., Ledeboer A. M., Venema G. Nucleotide sequence of the cell wall proteinase gene of Streptococcus cremoris Wg2. Appl Environ Microbiol. 1988 Jan;54(1):231–238. doi: 10.1128/aem.54.1.231-238.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Kondo J. K., McKay L. L. Plasmid transformation of Streptococcus lactis protoplasts: optimization and use in molecular cloning. Appl Environ Microbiol. 1984 Aug;48(2):252–259. doi: 10.1128/aem.48.2.252-259.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  17. Lakshmidevi G., Davidson B. E., Hillier A. J. Molecular characterization of promoters of the Lactococcus lactis subsp. cremoris temperate bacteriophage BK5-T and identification of a phage gene implicated in the regulation of promoter activity. Appl Environ Microbiol. 1990 Apr;56(4):934–942. doi: 10.1128/aem.56.4.934-942.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leenhouts K. J., Kok J., Venema G. Campbell-like integration of heterologous plasmid DNA into the chromosome of Lactococcus lactis subsp. lactis. Appl Environ Microbiol. 1989 Feb;55(2):394–400. doi: 10.1128/aem.55.2.394-400.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Maeda S., Gasson M. J. Cloning, expression and location of the Streptococcus lactis gene for phospho-beta-D-galactosidase. J Gen Microbiol. 1986 Feb;132(2):331–340. doi: 10.1099/00221287-132-2-331. [DOI] [PubMed] [Google Scholar]
  20. O'Callaghan C., Morris A. Inhibition of beta-lactamases by beta-lactam antibiotics. Antimicrob Agents Chemother. 1972 Dec;2(6):442–448. doi: 10.1128/aac.2.6.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Palva I. Molecular cloning of alpha-amylase gene from Bacillus amyloliquefaciens and its expression in B. subtilis. Gene. 1982 Jul-Aug;19(1):81–87. doi: 10.1016/0378-1119(82)90191-3. [DOI] [PubMed] [Google Scholar]
  22. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Scheirlinck T., Mahillon J., Joos H., Dhaese P., Michiels F. Integration and expression of alpha-amylase and endoglucanase genes in the Lactobacillus plantarum chromosome. Appl Environ Microbiol. 1989 Sep;55(9):2130–2137. doi: 10.1128/aem.55.9.2130-2137.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shaw W. V. Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria. Methods Enzymol. 1975;43:737–755. doi: 10.1016/0076-6879(75)43141-x. [DOI] [PubMed] [Google Scholar]
  25. Sibakov M., Koivula T., von Wright A., Palva I. Secretion of TEM beta-lactamase with signal sequences isolated from the chromosome of Lactococcus lactis subsp. lactis. Appl Environ Microbiol. 1991 Feb;57(2):341–348. doi: 10.1128/aem.57.2.341-348.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Simon D., Chopin A. Construction of a vector plasmid family and its use for molecular cloning in Streptococcus lactis. Biochimie. 1988 Apr;70(4):559–566. doi: 10.1016/0300-9084(88)90093-4. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Williams D. M., Duvall E. J., Lovett P. S. Cloning restriction fragments that promote expression of a gene in Bacillus subtilis. J Bacteriol. 1981 Jun;146(3):1162–1165. doi: 10.1128/jb.146.3.1162-1165.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. van de Guchte M., van der Vossen J. M., Kok J., Venema G. Construction of a lactococcal expression vector: expression of hen egg white lysozyme in Lactococcus lactis subsp. lactis. Appl Environ Microbiol. 1989 Jan;55(1):224–228. doi: 10.1128/aem.55.1.224-228.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. van der Vossen J. M., Kok J., Venema G. Construction of cloning, promoter-screening, and terminator-screening shuttle vectors for Bacillus subtilis and Streptococcus lactis. Appl Environ Microbiol. 1985 Aug;50(2):540–542. doi: 10.1128/aem.50.2.540-542.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. van der Vossen J. M., van der Lelie D., Venema G. Isolation and characterization of Streptococcus cremoris Wg2-specific promoters. Appl Environ Microbiol. 1987 Oct;53(10):2452–2457. doi: 10.1128/aem.53.10.2452-2457.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. von Wright A., Tynkkynen S., Suominen M. Cloning of a Streptococcus lactis subsp. lactis Chromosomal Fragment Associated with the Ability To Grow in Milk. Appl Environ Microbiol. 1987 Jul;53(7):1584–1588. doi: 10.1128/aem.53.7.1584-1588.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES