Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1993 Aug;59(8):2449–2456. doi: 10.1128/aem.59.8.2449-2456.1993

Effect of Increasing the Copy Number of Bacteriophage Origins of Replication, in trans, on Incoming-Phage Proliferation †

Daniel J O'Sullivan 1, Colin Hill 1,‡, Todd R Klaenhammer 1,*
PMCID: PMC182305  PMID: 16349010

Abstract

Bacteriophage resistance mechanisms which are derived from a bacteriophage genome are termed Per (phage-encoded resistance). When present in trans in Lactococcus lactis NCK203, Per50, the cloned origin of replication from phage φ50, interferes with φ50 replication. The per50 fragment was found to afford negligible protection to NCK203 against φ50 infection when present in a low-copy-number plasmid, pTRK325. A high-copy-number Per50 construct (pTRK323) dramatically affected φ50 infection, reducing the efficiency of plaquing (EOP) to 2.5 × 10-4 and the plaque size to pinhead proportions. This clone also afforded significant protection against other related small isometric phages. Per31 was cloned from phage φ31 and demonstrated to function as an origin of replication by enabling replication of per31-containing plasmids, in NCK203, on φ31 infection. A low-copy-number Per31 plasmid (pTRK360) reduced the EOP of φ31 on NCK203 to 0.3 and the plaque diameter from 1.5 to 0.5 mm. When this plasmid was cloned in high copy number, the EOP was further reduced to 7.2 × 10-7 but the plaques were large and contained Per31-resistant phages. Characterization of these “new” phages revealed at least two different types that were similar to φ31, except that DNA alterations were noted in the region containing the origin. This novel and powerful abortive phage resistance mechanism should prove useful when directed at specific, problematic phages.

Full text

PDF
2449

Images in this article

2453Image
on p.2453
2454Image
on p.2454
2454Image
on p.2454

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. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  3. Chopin A., Chopin M. C., Moillo-Batt A., Langella P. Two plasmid-determined restriction and modification systems in Streptococcus lactis. Plasmid. 1984 May;11(3):260–263. doi: 10.1016/0147-619x(84)90033-7. [DOI] [PubMed] [Google Scholar]
  4. Chung Y. B., Hinkle D. C. Bacteriophage T7 DNA packaging. I. Plasmids containing a T7 replication origin and the T7 concatemer junction are packaged into transducing particles during phage infection. J Mol Biol. 1990 Dec 20;216(4):911–926. doi: 10.1016/S0022-2836(99)80010-2. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Coffey A. G., Fitzgerald G. F., Daly C. Cloning and characterization of the determinant for abortive infection of bacteriophage from lactococcal plasmid pCI829. J Gen Microbiol. 1991 Jun;137(6):1355–1362. doi: 10.1099/00221287-137-6-1355. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Gautier M., Chopin M. C. Plasmid-Determined Systems for Restriction and Modification Activity and Abortive Infection in Streptococcus cremoris. Appl Environ Microbiol. 1987 May;53(5):923–927. doi: 10.1128/aem.53.5.923-927.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Harrington A., Hill C. Construction of a Bacteriophage-Resistant Derivative of Lactococcus lactis subsp. lactis 425A by Using the Conjugal Plasmid pNP40. Appl Environ Microbiol. 1991 Dec;57(12):3405–3409. doi: 10.1128/aem.57.12.3405-3409.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hill C., Miller L. A., Klaenhammer T. R. Cloning, expression, and sequence determination of a bacteriophage fragment encoding bacteriophage resistance in Lactococcus lactis. J Bacteriol. 1990 Nov;172(11):6419–6426. doi: 10.1128/jb.172.11.6419-6426.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hill C., Miller L. A., Klaenhammer T. R. Nucleotide sequence and distribution of the pTR2030 resistance determinant (hsp) which aborts bacteriophage infection in lactococci. Appl Environ Microbiol. 1990 Jul;56(7):2255–2258. doi: 10.1128/aem.56.7.2255-2258.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hill C., Pierce K., Klaenhammer T. R. The conjugative plasmid pTR2030 encodes two bacteriophage defense mechanisms in lactococci, restriction modification (R+/M+) and abortive infection (Hsp+). Appl Environ Microbiol. 1989 Sep;55(9):2416–2419. doi: 10.1128/aem.55.9.2416-2419.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jarvis A. W., Klaenhammer T. R. Bacteriophage Resistance Conferred on Lactic Streptococci by the Conjugative Plasmid pTR2030: Effects on Small Isometric-, Large Isometric-, and Prolate-Headed Phages. Appl Environ Microbiol. 1986 Jun;51(6):1272–1277. doi: 10.1128/aem.51.6.1272-1277.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Klaenhammer T. R., Sanozky R. B. Conjugal transfer from Streptococcus lactis ME2 of plasmids encoding phage resistance, nisin resistance and lactose-fermenting ability: evidence for a high-frequency conjugative plasmid responsible for abortive infection of virulent bacteriophage. J Gen Microbiol. 1985 Jun;131(6):1531–1541. doi: 10.1099/00221287-131-6-1531. [DOI] [PubMed] [Google Scholar]
  17. Krüger D. H., Bickle T. A. Bacteriophage survival: multiple mechanisms for avoiding the deoxyribonucleic acid restriction systems of their hosts. Microbiol Rev. 1983 Sep;47(3):345–360. doi: 10.1128/mr.47.3.345-360.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moineau S., Pandian S., Klaenhammer T. R. Restriction/Modification systems and restriction endonucleases are more effective on lactococcal bacteriophages that have emerged recently in the dairy industry. Appl Environ Microbiol. 1993 Jan;59(1):197–202. doi: 10.1128/aem.59.1.197-202.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'sullivan D. J., Klaenhammer T. R. Rapid Mini-Prep Isolation of High-Quality Plasmid DNA from Lactococcus and Lactobacillus spp. Appl Environ Microbiol. 1993 Aug;59(8):2730–2733. doi: 10.1128/aem.59.8.2730-2733.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Palca J. Errant HIV strain renders test virus stock useless. Science. 1992 Jun 5;256(5062):1387–1388. doi: 10.1126/science.1604310. [DOI] [PubMed] [Google Scholar]
  21. Powell I. B., Tulloch D. L., Hillier A. J., Davidson B. E. Phage DNA synthesis and host DNA degradation in the life cycle of Lactococcus lactis bacteriophage c6A. J Gen Microbiol. 1992 May;138(5):945–950. doi: 10.1099/00221287-138-5-945. [DOI] [PubMed] [Google Scholar]
  22. Sanders M. E., Klaenhammer T. R. Characterization of Phage-Sensitive Mutants from a Phage-Insensitive Strain of Streptococcus lactis: Evidence for a Plasmid Determinant that Prevents Phage Adsorption. Appl Environ Microbiol. 1983 Nov;46(5):1125–1133. doi: 10.1128/aem.46.5.1125-1133.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Sanders M. E., Leonhard P. J., Sing W. D., Klaenhammer T. R. Conjugal strategy for construction of fast Acid-producing, bacteriophage-resistant lactic streptococci for use in dairy fermentations. Appl Environ Microbiol. 1986 Nov;52(5):1001–1007. doi: 10.1128/aem.52.5.1001-1007.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Sing W. D., Klaenhammer T. R. Conjugal Transfer of Bacteriophage Resistance Determinants on pTR2030 into Streptococcus cremoris Strains. Appl Environ Microbiol. 1986 Jun;51(6):1264–1271. doi: 10.1128/aem.51.6.1264-1271.1986. [DOI] [PMC free article] [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]

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

RESOURCES