Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Sep;163(3):1229–1236. doi: 10.1128/jb.163.3.1229-1236.1985

N conjugative transfer system of plasmid pCU1.

V Thatte, D E Bradley, V N Iyer
PMCID: PMC219264  PMID: 2863255

Abstract

The mating and plasmid DNA transfer functions in Escherichia coli K-12 strains that are determined by the IncN group plasmid pCU1 are specified by a single 19.4-kilobase (kb) segment of the 39-kb plasmid. Analysis of this tra region by transposon Tn5 mutagenesis and by complementation tests indicated that a subset of tra comprising six complementation groups (presumably transcription units) determines sensitivity to the N pilus-specific bacteriophages IKe and PRD1. Deletion derivatives and molecular clones that contained only this part of tra conferred sensitivity to IKe and PRD1 and the ability to produce N pili without conferring the Tra phenotype. The entire tra region contained two other complementation groups. If the 19.4-kb tra region is interrupted by DNA that is irrelevant to the Tra phenotype, such DNA must be less than 2 kb long. There are two regions within tra where this is a possibility. These observations are compared with those obtained recently with a related plasmid.

Full text

PDF
1229

Images in this article

1232Image
on p.1232
1232Image
on p.1232
1234Image
on p.1234

Selected References

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

  1. Barth P. T., Grinter N. J., Bradley D. E. Conjugal transfer system of plasmid RP4: analysis by transposon 7 insertion. J Bacteriol. 1978 Jan;133(1):43–52. doi: 10.1128/jb.133.1.43-52.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bradley D. E. Morphological and serological relationships of conjugative pili. Plasmid. 1980 Sep;4(2):155–169. doi: 10.1016/0147-619x(80)90005-0. [DOI] [PubMed] [Google Scholar]
  4. Bradley D. E. Morphology of pili determined by the N incompatibility group plasmid N3 and interaction with bacteriophages PR4 and IKe. Plasmid. 1979 Oct;2(4):632–636. doi: 10.1016/0147-619x(79)90061-1. [DOI] [PubMed] [Google Scholar]
  5. Brown A. M., Willetts N. S. A physical and genetic map of the IncN plasmid R46. Plasmid. 1981 Mar;5(2):188–201. doi: 10.1016/0147-619x(81)90020-2. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. 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]
  9. Dennison S., Baumberg S. Conjugational behaviour of N plasmids in Escherichia coli K12. Mol Gen Genet. 1975 Jul 10;138(4):323–331. doi: 10.1007/BF00264802. [DOI] [PubMed] [Google Scholar]
  10. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  11. Humphreys G. O., Willshaw G. A., Anderson E. S. A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta. 1975 Apr 2;383(4):457–463. doi: 10.1016/0005-2787(75)90318-4. [DOI] [PubMed] [Google Scholar]
  12. Kahn M., Kolter R., Thomas C., Figurski D., Meyer R., Remaut E., Helinski D. R. Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol. 1979;68:268–280. doi: 10.1016/0076-6879(79)68019-9. [DOI] [PubMed] [Google Scholar]
  13. Khatoon H., Iyer R. V., Iyer V. N. A new filamentous bacteriophage with sex-factor specificity. Virology. 1972 Apr;48(1):145–155. doi: 10.1016/0042-6822(72)90122-5. [DOI] [PubMed] [Google Scholar]
  14. Konarska-Kozlowska M., Thatte V., Iyer V. N. Inverted repeats in the DNA of plasmid pCU1. J Bacteriol. 1983 Mar;153(3):1502–1512. doi: 10.1128/jb.153.3.1502-1512.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Langer P. J., Walker G. C. Restriction endonuclease cleavage map of pKM101: relationship to parental plasmid R46. Mol Gen Genet. 1981;182(2):268–272. doi: 10.1007/BF00269669. [DOI] [PubMed] [Google Scholar]
  16. Murray K., Murray N. E. Phage lambda receptor chromosomes for DNA fragments made with restriction endonuclease III of Haemophilus influenzae and restriction endonuclease I of Escherichia coli. J Mol Biol. 1975 Nov 5;98(3):551–564. doi: 10.1016/s0022-2836(75)80086-6. [DOI] [PubMed] [Google Scholar]
  17. Olsen R. H., Siak J. S., Gray R. H. Characteristics of PRD1, a plasmid-dependent broad host range DNA bacteriophage. J Virol. 1974 Sep;14(3):689–699. doi: 10.1128/jvi.14.3.689-699.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rodriguez M., Iyer V. N. Killing of Klebsiella pneumoniae mediated by conjugation with bacteria carrying antibiotic-resistance plasmids of the group N. Plasmid. 1981 Jul;6(1):141–147. doi: 10.1016/0147-619x(81)90060-3. [DOI] [PubMed] [Google Scholar]
  19. Selvaraj G., Iyer V. N. Suicide plasmid vehicles for insertion mutagenesis in Rhizobium meliloti and related bacteria. J Bacteriol. 1983 Dec;156(3):1292–1300. doi: 10.1128/jb.156.3.1292-1300.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tait R. C., Lundquist R. C., Kado C. I. Genetic map of the crown gall suppressive IncW plasmid pSa. Mol Gen Genet. 1982;186(1):10–15. doi: 10.1007/BF00422905. [DOI] [PubMed] [Google Scholar]
  21. Thatte V., Gill S., Iyer V. N. Regions on plasmid pCU1 required for the killing of Klebsiella pneumoniae. J Bacteriol. 1985 Sep;163(3):1296–1299. doi: 10.1128/jb.163.3.1296-1299.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thatte V., Iyer V. N. Cloning of a plasmid region specifying the N transfer system of bacterial conjugation in Escherichia coli. Gene. 1983 Mar;21(3):227–236. doi: 10.1016/0378-1119(83)90006-9. [DOI] [PubMed] [Google Scholar]
  23. Ward J. M., Grinsted J. Physical and genetic analysis of the Inc-W group plasmids R388, Sa, and R7K. Plasmid. 1982 May;7(3):239–250. doi: 10.1016/0147-619x(82)90005-1. [DOI] [PubMed] [Google Scholar]
  24. Winans S. C., Walker G. C. Entry exclusion determinant(s) of IncN plasmid pKM101. J Bacteriol. 1985 Jan;161(1):411–416. doi: 10.1128/jb.161.1.411-416.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Winans S. C., Walker G. C. Genetic localization and characterization of a pKM101-coded endonuclease. J Bacteriol. 1983 Jun;154(3):1117–1125. doi: 10.1128/jb.154.3.1117-1125.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES