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. 1981 Aug;78(8):4902–4906. doi: 10.1073/pnas.78.8.4902

Replication of plasmid pT181 DNA in vitro: requirement for a plasmid-encoded product.

S A Khan, S M Carleton, R P Novick
PMCID: PMC320289  PMID: 6946436

Abstract

PT181 is a naturally occurring 4.5-kilobase Staphylococcus aureus plasmid encoding resistance to tetracycline. The plasmid has a copy number of about 20 per cell; a mutant, cop-608, that has a copy number of 800-1000 has been isolated. A cell-free extract has been developed that carries out complete replication of this plasmid. Extracts made from a strain containing the mutant have much greater replication activity than do extracts of strains containing pT181. In an extract from which endogenous DNA has been removed, DNA synthesis is dependent upon the addition of exogenous plasmid DNA. The replication system is specific for pT181 and related plasmids but it is inactive with other S. aureus plasmids. Furthermore, pT181 DNA does not replicate in extracts made from plasmid-negative strains or strains containing other plasmids. The results suggest that a specific plasmid-encoded substance is required for the replication of pT181 DNA.

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Selected References

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

  1. Bezanson G. S., Goebel W. In vitro system for the replication of the mini R1 factor Rsc11. Mol Gen Genet. 1979 Feb 16;170(1):49–56. doi: 10.1007/BF00268579. [DOI] [PubMed] [Google Scholar]
  2. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Conrad S. E., Campbell J. L. Characterization of an improved in vitro DNA replication system for Escherichia coli plasmids. Nucleic Acids Res. 1979 Jul 25;6(10):3289–3304. doi: 10.1093/nar/6.10.3289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Donoghue D. J., Sharp P. A. Replication of colicin E1 plasmid DNA in vivo requires no plasmid-encoded proteins. J Bacteriol. 1978 Mar;133(3):1287–1294. doi: 10.1128/jb.133.3.1287-1294.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Inuzuka M., Helinski D. R. Replication of antibiotic resistance plasmid R6K DNA in vitro. Biochemistry. 1978 Jun 27;17(13):2567–2573. doi: 10.1021/bi00606a017. [DOI] [PubMed] [Google Scholar]
  6. Inuzuka M., Helinski D. R. Requirement of a plasmid-encoded protein for replication in vitro of plasmid R6K. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5381–5385. doi: 10.1073/pnas.75.11.5381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Iordanescu S., Surdeanu M., Della Latta P., Novick R. Incompatibility and molecular relationships between small Staphylococcal plasmids carrying the same resistance marker. Plasmid. 1978 Sep;1(4):468–479. doi: 10.1016/0147-619x(78)90005-7. [DOI] [PubMed] [Google Scholar]
  8. Iordănescu S. Temperature-sensitive mutant of a tetracycline resistance staphylococcal plasmid. Arch Roum Pathol Exp Microbiol. 1976 Jul;35(3):257–264. [PubMed] [Google Scholar]
  9. Kolter R., Inuzuka M., Figurski D., Thomas C., Stalker D., Helinski D. R. Plasmid DNA replication: RK2- and R6K-encoded trans-acting factors and their sites of action. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):91–97. doi: 10.1101/sqb.1979.043.01.014. [DOI] [PubMed] [Google Scholar]
  10. Kreuzer K. N., Cozzarelli N. R. Formation and resolution of DNA catenanes by DNA gyrase. Cell. 1980 May;20(1):245–254. doi: 10.1016/0092-8674(80)90252-4. [DOI] [PubMed] [Google Scholar]
  11. NOVICK R. P. ANALYSIS BY TRANSDUCTION OF MUTATIONS AFFECTING PENICILLINASE FORMATION IN STAPHYLOCOCCUS AUREUS. J Gen Microbiol. 1963 Oct;33:121–136. doi: 10.1099/00221287-33-1-121. [DOI] [PubMed] [Google Scholar]
  12. Nordström K., Ingram L. C., Lundbäck A. Mutations in R factors of Escherichia coli causing an increased number of R-factor copies per chromosome. J Bacteriol. 1972 May;110(2):562–569. doi: 10.1128/jb.110.2.562-569.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Novick R. P., Brodsky R. Studies on plasmid replication. I. Plasmid incompatibility and establishment in Staphylococcus aureus. J Mol Biol. 1972 Jul 21;68(2):285–302. doi: 10.1016/0022-2836(72)90214-8. [DOI] [PubMed] [Google Scholar]
  14. Novick R. P., Murphy E., Gryczan T. J., Baron E., Edelman I. Penicillinase plasmids of Staphylococcus aureus: restriction-deletion maps. Plasmid. 1979 Jan;2(1):109–129. doi: 10.1016/0147-619x(79)90010-6. [DOI] [PubMed] [Google Scholar]
  15. Novick R. P. Studies on plasmid replication. III. Isolation and characterization of replication-defective mutants. Mol Gen Genet. 1974;135(2):131–147. doi: 10.1007/BF00264781. [DOI] [PubMed] [Google Scholar]
  16. Sakakibara Y., Tomizawa J. I. Replication of colicin E1 plasmid DNA in cell extracts. Proc Natl Acad Sci U S A. 1974 Mar;71(3):802–806. doi: 10.1073/pnas.71.3.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sakakibara Y., Tomizawa J. Replication of colicin E1 plasmid DNA in cell extracts. II. Selective synthesis of early replicative intermediates. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1403–1407. doi: 10.1073/pnas.71.4.1403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shalita Z., Hertman I., Sarid S. Isolation and characterization of a plasmid involved with enterotoxin B production in Staphylococcus aureus. J Bacteriol. 1977 Jan;129(1):317–325. doi: 10.1128/jb.129.1.317-325.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Staudenbauer W. L. Replication of small plasmids in extracts of Escherichia coli. Mol Gen Genet. 1976 Jun 15;145(3):273–280. doi: 10.1007/BF00325823. [DOI] [PubMed] [Google Scholar]
  20. Tomizawa J. I., Sakakibara Y., Kakefuda T. Replication of colicin E1 plasmid DNA added to cell extracts. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1050–1054. doi: 10.1073/pnas.72.3.1050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wyman L., Novick R. P. Studies on plasmid replication. IV. Complementation of replication-defective mutants by an incompatibility-deficient plasmid. Mol Gen Genet. 1974;135(2):149–161. doi: 10.1007/BF00264782. [DOI] [PubMed] [Google Scholar]

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