Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1977 Mar;74(3):1138–1141. doi: 10.1073/pnas.74.3.1138

Bidirection replication from a unique origin in a mini-F plasmid.

R Eichenlaub, D Figurski, D R Helinski
PMCID: PMC430623  PMID: 322144

Abstract

Replicating molecules of the mini F-kanamycin resistance plasmid, pML31, derived from F'lac, have been isolated from Escherichia coli as covalently-closed circular DNA molecules. These molecules were examined in the electron microscope after digestion with either EcoRI or BamHI restriction endonuclease. The structure of the majority of the molecules replicating was consistent with a bidirectional mode of replication starting at a unique origin on the F-fragment. This origin is located approximately 2.3 kilobases from one of the EcoRI sites. Orientation of the F-fragment relative to the physical map of F showed the position of the origin to be at 42.6 kilobases. A small proportion of molecules appeared to be replicating unidirectionally in either direction from this origin. Termination of replication of pML31 apparently occurs in the fragment containing the locus for kanamycin resistance in a unique region opposite the origin in the circular DNA molecule.

Full text

PDF
1139

Images in this article

Selected References

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

  1. Bazaral M., Helinski D. R. Characterization of multiple circular DNA forms of colicinogenic factor E-1 from Proteus mirabilis. Biochemistry. 1968 Oct;7(10):3513–3520. doi: 10.1021/bi00850a028. [DOI] [PubMed] [Google Scholar]
  2. Bourgaux P., Bourgaux-Ramoisy D. Unwinding of replicating polyoma virus DNA. J Mol Biol. 1972 Oct 14;70(3):399–413. doi: 10.1016/0022-2836(72)90548-7. [DOI] [PubMed] [Google Scholar]
  3. Cabello F., Timmis K., Cohen S. N. Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature. 1976 Jan 29;259(5541):285–290. doi: 10.1038/259285a0. [DOI] [PubMed] [Google Scholar]
  4. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Guerry P., LeBlanc D. J., Falkow S. General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973 Nov;116(2):1064–1066. doi: 10.1128/jb.116.2.1064-1066.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guyer M. S., Figurski D., Davidson N. Electron microscope study of a plasmid chimera containing the replication region of the Escherichia coli F plasmid. J Bacteriol. 1976 Aug;127(2):988–997. doi: 10.1128/jb.127.2.988-997.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gyurasits E. B., Wake R. G. Bidirectional chromosome replication in Bacillus subtilis. J Mol Biol. 1973 Jan;73(1):55–63. doi: 10.1016/0022-2836(73)90158-7. [DOI] [PubMed] [Google Scholar]
  8. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jaenisch R., Mayer A., Levine A. Replicating SV40 molecules containing closed circular template DNA strands. Nat New Biol. 1971 Sep 15;233(37):72–75. doi: 10.1038/newbio233072a0. [DOI] [PubMed] [Google Scholar]
  10. Jaenisch R., Mayer A., Levine A. Replicating SV40 molecules containing closed circular template DNA strands. Nat New Biol. 1971 Sep 15;233(37):72–75. doi: 10.1038/newbio233072a0. [DOI] [PubMed] [Google Scholar]
  11. Lovett M. A., Helinski D. R. Method for the isolation of the replication region of a bacterial replicon: construction of a mini-F'kn plasmid. J Bacteriol. 1976 Aug;127(2):982–987. doi: 10.1128/jb.127.2.982-987.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lovett M. A., Katz L., Helinski D. R. Unidirectional replication of plasmid ColE1 DNA. Nature. 1974 Sep 27;251(5473):337–340. doi: 10.1038/251337a0. [DOI] [PubMed] [Google Scholar]
  13. Lovett M. A., Sparks R. B., Helinski D. R. Bidirectional replication of plasmid R6K DNA in Escherichia coli; correspondence between origin of replication and position of single-strand break in relaxed complex. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2905–2909. doi: 10.1073/pnas.72.8.2905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Masters M., Broda P. Evidence for the bidirectional replications of the Escherichia coli chromosome. Nat New Biol. 1971 Aug 4;232(31):137–140. doi: 10.1038/newbio232137a0. [DOI] [PubMed] [Google Scholar]
  15. Nishioka Y., Eisenstark A. Sequence of genes replicated in Salmonella typhimurium as examined by transduction techniques. J Bacteriol. 1970 May;102(2):320–333. doi: 10.1128/jb.102.2.320-333.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Palchaudhuri S., Maas W. K. Physical mapping of a DNA sequence common to plasmids of incompatibility group F I. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1190–1194. doi: 10.1073/pnas.74.3.1190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schnös M., Inman R. B. Position of branch points in replicating lambda DNA. J Mol Biol. 1970 Jul 14;51(1):61–73. doi: 10.1016/0022-2836(70)90270-6. [DOI] [PubMed] [Google Scholar]
  18. Sebring E. D., Kelly T. J., Jr, Thoren M. M., Salzman N. P. Structure of replicating simian virus 40 deoxyribonucleic acid molecules. J Virol. 1971 Oct;8(4):478–490. doi: 10.1128/jvi.8.4.478-490.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sharp P. A., Hsu M. T., Otsubo E., Davidson N. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. I. Structure of F-prime factors. J Mol Biol. 1972 Nov 14;71(2):471–497. doi: 10.1016/0022-2836(72)90363-4. [DOI] [PubMed] [Google Scholar]
  20. Wilson G. A., Young F. E. Isolation of a sequence-specific endonuclease (BamI) from Bacillus amyloliquefaciens H. J Mol Biol. 1975 Sep 5;97(1):123–125. doi: 10.1016/s0022-2836(75)80028-3. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES