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. 1994 Feb;176(4):1069–1076. doi: 10.1128/jb.176.4.1069-1076.1994

Characterization of the lactococcal temperate phage TP901-1 and its site-specific integration.

B Christiansen 1, M G Johnsen 1, E Stenby 1, F K Vogensen 1, K Hammer 1
PMCID: PMC205158  PMID: 8106318

Abstract

The temperate lactococcal phage TP901-1, induced by UV light from Lactococcus lactis subsp. cremoris 901-1, was characterized. The restriction map was found to be circular, and the packaging of TP901-1 DNA was concluded to occur by a headful mechanism. The pac region was localized on the 38.4-kb phage genome. TP901-1 belongs to the class of P335 phages (V. Braun, S. Hertwig, H. Neve, A. Geis, and M. Teuber, J. Gen. Microbiol. 135:2551-2560, 1989). Evidence is presented that the phages TP936-1 (V. Braun, S. Hertwig, H. Neve, A. Geis, and M. Teuber, J. Gen. Microbiol. 135:2551-2560, 1989) and C3-T1 (A. W. Jarvis, V. R. Parker, and M. B. Bianchin, Can. J. Microbiol. 38:398-404, 1992) are very closely related to or are identical to TP901-1. The lytically propagated TP901-1 phages were able to lysogenize both indicator strains Lactococcus cremoris 3107 and Wg2. Lysogenization resulted in site-specific integration of the phage genome into the bacterial chromosome. Only one chromosomal attB site was found in 20 independent lysogens. The attP region of TP901-1 and the attL and attR regions were cloned and sequenced. The results showed a core region of only 5 bp, in which the recombination occurs, followed after a 1-bp mismatch by a 7-bp identical region, TCAAT(T/C)AAGGTAA. This result was further verified by sequencing of the attB region obtained by PCR. An integration vector was constructed with the 6.5-kb EcoRI fragment from TP901-1 containing attP. This vector also functions in the plasmid-free strains, MG1363 and LM0230 with only one specific attB site, strongly indicating a more general use of the TP901-1-based integration vector in lactococci.

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Selected References

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  1. Campbell A. M. Chromosomal insertion sites for phages and plasmids. J Bacteriol. 1992 Dec;174(23):7495–7499. doi: 10.1128/jb.174.23.7495-7499.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Casjens S., Huang W. M., Hayden M., Parr R. Initiation of bacteriophage P22 DNA packaging series. Analysis of a mutant that alters the DNA target specificity of the packaging apparatus. J Mol Biol. 1987 Apr 5;194(3):411–422. doi: 10.1016/0022-2836(87)90671-1. [DOI] [PubMed] [Google Scholar]
  3. Coleman D., Knights J., Russell R., Shanley D., Birkbeck T. H., Dougan G., Charles I. Insertional inactivation of the Staphylococcus aureus beta-toxin by bacteriophage phi 13 occurs by site- and orientation-specific integration of the phi 13 genome. Mol Microbiol. 1991 Apr;5(4):933–939. doi: 10.1111/j.1365-2958.1991.tb00768.x. [DOI] [PubMed] [Google Scholar]
  4. Efstathiou J. D., McKay L. L. Inorganic salts resistance associated with a lactose-fermenting plasmid in Streptococcus lactis. J Bacteriol. 1977 Apr;130(1):257–265. doi: 10.1128/jb.130.1.257-265.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gasson M. J., Davies F. L. Prophage-Cured Derivatives of Streptococcus lactis and Streptococcus cremoris. Appl Environ Microbiol. 1980 Nov;40(5):964–966. doi: 10.1128/aem.40.5.964-966.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Hauser M. A., Scocca J. J. Location of the host attachment site for phage HPl within a cluster of Haemophilus influenzae tRNA genes. Nucleic Acids Res. 1990 Sep 11;18(17):5305–5305. doi: 10.1093/nar/18.17.5305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Lakshmidevi G., Davidson B. E., Hillier A. J. Circular Permutation of the Genome of a Temperate Bacteriophage from Streptococcus cremoris BK5. Appl Environ Microbiol. 1988 Apr;54(4):1039–1045. doi: 10.1128/aem.54.4.1039-1045.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Landy A., Ross W. Viral integration and excision: structure of the lambda att sites. Science. 1977 Sep 16;197(4309):1147–1160. doi: 10.1126/science.331474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lee C. Y., Iandolo J. J. Integration of staphylococcal phage L54a occurs by site-specific recombination: structural analysis of the attachment sites. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5474–5478. doi: 10.1073/pnas.83.15.5474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lee C. Y., Iandolo J. J. Structural analysis of staphylococcal bacteriophage phi 11 attachment sites. J Bacteriol. 1988 May;170(5):2409–2411. doi: 10.1128/jb.170.5.2409-2411.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lee M. H., Pascopella L., Jacobs W. R., Jr, Hatfull G. F. Site-specific integration of mycobacteriophage L5: integration-proficient vectors for Mycobacterium smegmatis, Mycobacterium tuberculosis, and bacille Calmette-Guérin. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3111–3115. doi: 10.1073/pnas.88.8.3111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Leong J. M., Nunes-Düby S. E., Oser A. B., Lesser C. F., Youderian P., Susskind M. M., Landy A. Structural and regulatory divergence among site-specific recombination genes of lambdoid phage. J Mol Biol. 1986 Jun 20;189(4):603–616. doi: 10.1016/0022-2836(86)90491-2. [DOI] [PubMed] [Google Scholar]
  16. Lillehaug D., Birkeland N. K. Characterization of genetic elements required for site-specific integration of the temperate lactococcal bacteriophage phi LC3 and construction of integration-negative phi LC3 mutants. J Bacteriol. 1993 Mar;175(6):1745–1755. doi: 10.1128/jb.175.6.1745-1755.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lillehaug D., Lindqvist B., Birkeland N. K. Characterization of phiLC3, a Lactococcus lactis subsp. cremoris temperature bacteriophage with cohesive single-stranded DNA ends. Appl Environ Microbiol. 1991 Nov;57(11):3206–3211. doi: 10.1128/aem.57.11.3206-3211.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Raya R. R., Fremaux C., De Antoni G. L., Klaenhammer T. R. Site-specific integration of the temperate bacteriophage phi adh into the Lactobacillus gasseri chromosome and molecular characterization of the phage (attP) and bacterial (attB) attachment sites. J Bacteriol. 1992 Sep;174(17):5584–5592. doi: 10.1128/jb.174.17.5584-5592.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Reiter W. D., Palm P., Yeats S. Transfer RNA genes frequently serve as integration sites for prokaryotic genetic elements. Nucleic Acids Res. 1989 Mar 11;17(5):1907–1914. doi: 10.1093/nar/17.5.1907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. 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]
  23. Waldman A. S., Goodman S. D., Scocca J. J. Nucleotide sequences and properties of the sites involved in lysogenic insertion of the bacteriophage HP1c1 genome into the Haemophilus influenzae chromosome. J Bacteriol. 1987 Jan;169(1):238–246. doi: 10.1128/jb.169.1.238-246.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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