Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Sep;62(9):3075–3082. doi: 10.1128/aem.62.9.3075-3082.1996

AbiG, a genotypically novel abortive infection mechanism encoded by plasmid pCI750 of Lactococcus lactis subsp. cremoris UC653.

L O'Connor 1, A Coffey 1, C Daly 1, G F Fitzgerald 1
PMCID: PMC168098  PMID: 8795193

Abstract

AbiG is an abortive infection (Abi) mechanism encoded by the conjugative plasmid pCI750 originally isolated from Lactococcus lactis subsp. cremoris UC653. Insensitivity conferred by this Abi manifested itself as complete resistance to phi 712 (936 phage species) with only partial resistance to phi c2 (c2 species). The mechanism did not inhibit phage DNA replication. The smallest subclone of pCI750 which expressed the Abi phenotype contained a 3.5-kb insert which encoded two potential open reading frames. abiGi (750 bp) and abiGii (1,194 bp) were separated by 2 bp and appeared to share a single promoter upstream of abiGi. These open reading frames showed no significant homology to sequences of either the DNA or protein databases; however, they did exhibit the typical low G+C content (29 and 27%, respectively) characteristic of lactococcal abi genes. In fact, the G+C content of a 7.0-kb fragment incorporating the abiG locus was 30%, which may suggest horizontal gene transfer from a species of low G+C content. In this context, it is notable that remnants of IS elements were observed throughout this 7.0-kb region.

Full Text

The Full Text of this article is available as a PDF (454.4 KB).

Selected References

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

  1. Alatossava T., Klaenhammer T. R. Molecular Characterization of Three Small Isometric-Headed Bacteriophages Which Vary in Their Sensitivity to the Lactococcal Phage Resistance Plasmid pTR2030. Appl Environ Microbiol. 1991 May;57(5):1346–1353. doi: 10.1128/aem.57.5.1346-1353.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Anba J., Bidnenko E., Hillier A., Ehrlich D., Chopin M. C. Characterization of the lactococcal abiD1 gene coding for phage abortive infection. J Bacteriol. 1995 Jul;177(13):3818–3823. doi: 10.1128/jb.177.13.3818-3823.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Behnke D., Gilmore M. S., Ferretti J. J. Plasmid pGB301, a new multiple resistance streptococcal cloning vehicle and its use in cloning of a gentamicin/kanamycin resistance determinant. Mol Gen Genet. 1981;182(3):414–421. doi: 10.1007/BF00293929. [DOI] [PubMed] [Google Scholar]
  6. Behnke D., Malke H. Bacteriophage interference in Streptococcus pyogenes. I. Characterization of prophage--host systems interfering with the virulent phage A25. Virology. 1978 Mar;85(1):118–128. doi: 10.1016/0042-6822(78)90416-6. [DOI] [PubMed] [Google Scholar]
  7. Bidnenko E., Ehrlich D., Chopin M. C. Phage operon involved in sensitivity to the Lactococcus lactis abortive infection mechanism AbiD1. J Bacteriol. 1995 Jul;177(13):3824–3829. doi: 10.1128/jb.177.13.3824-3829.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Chopin A. Organization and regulation of genes for amino acid biosynthesis in lactic acid bacteria. FEMS Microbiol Rev. 1993 Sep;12(1-3):21–37. doi: 10.1111/j.1574-6976.1993.tb00011.x. [DOI] [PubMed] [Google Scholar]
  11. Cluzel P. J., Chopin A., Ehrlich S. D., Chopin M. C. Phage abortive infection mechanism from Lactococcus lactis subsp. lactis, expression of which is mediated by an Iso-ISS1 element. Appl Environ Microbiol. 1991 Dec;57(12):3547–3551. doi: 10.1128/aem.57.12.3547-3551.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Coveney J. A., Fitzgerald G. F., Daly C. Detailed characterization and comparison of four lactic streptococcal bacteriophages based on morphology, restriction mapping, DNA homology, and structural protein analysis. Appl Environ Microbiol. 1987 Jul;53(7):1439–1447. doi: 10.1128/aem.53.7.1439-1447.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dao M. L., Ferretti J. J. Streptococcus-Escherichia coli shuttle vector pSA3 and its use in the cloning of streptococcal genes. Appl Environ Microbiol. 1985 Jan;49(1):115–119. doi: 10.1128/aem.49.1.115-119.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Durmaz E., Higgins D. L., Klaenhammer T. R. Molecular characterization of a second abortive phage resistance gene present in Lactococcus lactis subsp. lactis ME2. J Bacteriol. 1992 Nov;174(22):7463–7469. doi: 10.1128/jb.174.22.7463-7469.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Durmaz E., Klaenhammer T. R. A Starter Culture Rotation Strategy Incorporating Paired Restriction/ Modification and Abortive Infection Bacteriophage Defenses in a Single Lactococcus lactis Strain. Appl Environ Microbiol. 1995 Apr;61(4):1266–1273. doi: 10.1128/aem.61.4.1266-1273.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Garvey P., Hill C., Fitzgerald G. F. The Lactococcal Plasmid pNP40 Encodes a Third Bacteriophage Resistance Mechanism, One Which Affects Phage DNA Penetration. Appl Environ Microbiol. 1996 Feb;62(2):676–679. doi: 10.1128/aem.62.2.676-679.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grkovic S., O'callaghan M., Mahanty H. K. Characterization of Serratia entomophila Bacteriophages and the Phage-Resistant Mutant Strain BC4B. Appl Environ Microbiol. 1995 Dec;61(12):4160–4166. doi: 10.1128/aem.61.12.4160-4166.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Harley C. B., Reynolds R. P. Analysis of E. coli promoter sequences. Nucleic Acids Res. 1987 Mar 11;15(5):2343–2361. doi: 10.1093/nar/15.5.2343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hayes F., Daly C., Fitzgerald G. F. Identification of the Minimal Replicon of Lactococcus lactis subsp. lactis UC317 Plasmid pCI305. Appl Environ Microbiol. 1990 Jan;56(1):202–209. doi: 10.1128/aem.56.1.202-209.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hill C., Massey I. J., Klaenhammer T. R. Rapid method to characterize lactococcal bacteriophage genomes. Appl Environ Microbiol. 1991 Jan;57(1):283–288. doi: 10.1128/aem.57.1.283-288.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hill C., Miller L. A., Klaenhammer T. R. In vivo genetic exchange of a functional domain from a type II A methylase between lactococcal plasmid pTR2030 and a virulent bacteriophage. J Bacteriol. 1991 Jul;173(14):4363–4370. doi: 10.1128/jb.173.14.4363-4370.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  26. McLandsborough L. A., Kolaetis K. M., Requena T., McKay L. L. Cloning and characterization of the abortive infection genetic determinant abiD isolated from pBF61 of Lactococcus lactis subsp. lactis KR5. Appl Environ Microbiol. 1995 May;61(5):2023–2026. doi: 10.1128/aem.61.5.2023-2026.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moineau S., Pandian S., Klaenhammer T. R. Evolution of a Lytic Bacteriophage via DNA Acquisition from the Lactococcus lactis Chromosome. Appl Environ Microbiol. 1994 Jun;60(6):1832–1841. doi: 10.1128/aem.60.6.1832-1841.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Polzin K. M., McKay L. L. Identification, DNA sequence, and distribution of IS981, a new, high-copy-number insertion sequence in lactococci. Appl Environ Microbiol. 1991 Mar;57(3):734–743. doi: 10.1128/aem.57.3.734-743.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Polzin K. M., Shimizu-Kadota M. Identification of a new insertion element, similar to gram-negative IS26, on the lactose plasmid of Streptococcus lactis ML3. J Bacteriol. 1987 Dec;169(12):5481–5488. doi: 10.1128/jb.169.12.5481-5488.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Romero D. A., Klaenhammer T. R. Characterization of insertion sequence IS946, an Iso-ISS1 element, isolated from the conjugative lactococcal plasmid pTR2030. J Bacteriol. 1990 Aug;172(8):4151–4160. doi: 10.1128/jb.172.8.4151-4160.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sanders M. E., Klaenhammer T. R. Restriction and modification in group N streptococci: effect of heat on development of modified lytic bacteriophage. Appl Environ Microbiol. 1980 Sep;40(3):500–506. doi: 10.1128/aem.40.3.500-506.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Waterfield N. R., Le Page R. W., Wilson P. W., Wells J. M. The isolation of lactococcal promoters and their use in investigating bacterial luciferase synthesis in Lactococcus lactis. Gene. 1995 Nov 7;165(1):9–15. doi: 10.1016/0378-1119(95)00484-n. [DOI] [PubMed] [Google Scholar]
  35. Wirth R., An F. Y., Clewell D. B. Highly efficient protoplast transformation system for Streptococcus faecalis and a new Escherichia coli-S. faecalis shuttle vector. J Bacteriol. 1986 Mar;165(3):831–836. doi: 10.1128/jb.165.3.831-836.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  37. 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]

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

RESOURCES