Skip to main content
SpringerLink
Log in

Deletion mapping of kil and kor functions in the trfA and trfB regions of broad host range plasmid RK2

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Figurski et al. (1982) have reported that certain loci on the broad host range plasmid RK2 (kil functions) can be cloned only in the presence of other trans-acting segments of the plasmid genome (kor functions). They have suggested that the presence of these functions may in part account for the structure of mini RK2 replicons which were constructed in order to define the regions of the plasmid which encode replication/maintenance functions (Thomas et al. 1980). We have therefore investigated the relationship between these two sets of kil and kor loci and the loci implicated in the replication/maintenance of RK2. We find that, whilst the three kil loci reported by Figurski et al. (1982) are absent from these derivatives, a fourth such locus (kilD) is closely linked to trfA, a gene essential for RK2 replication. The kilD locus was probably responsible for the inclusion in mini replicons of a segment of PK2 DNA which carries both korD and korA in addition to trfB, a gene defined by a temperature-sensitive maintenance defect, but which can be deleted leaving a functional RK2 replicon (Thomas 1981 b). The kilB locus is situated on the opposite side of kilD from trfA, all three loci lying within a 3.6 kb segment of RK2 DNA. The korA, korD and trfB functions all map within a 900 bp segment of DNA, while korB requires sequence information at least 1.5 kb from this segment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Barth PT (1979) RP4 and R300B as wide host-range cloning vehicles. In: Timmis KN, Pühler A (eds) Plamids of medical, environmental and commercial importance. Elsevier/North-Holland, Amsterdam, p 399–410

    Google Scholar 

  • Beringer JE (1974) R factor transfer in Rhizobium leguminosarum. J Gen Microbiol 84:188–198

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7:1513–1523

    Google Scholar 

  • Bolivar R, Rodriguez RL, Green PJ, Betlach MC, Heynecker HC, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113

    Google Scholar 

  • Burkhardt HJ, Riess G, Pühler A (1979) Relationship of group P1 plasmids revealed by heteroduplex experiments: RP1, RP4, R68 and RK2 are identical. J Gen Microbiol 114:341–348

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic mini plasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Cho JJ, Panopoulos NJ, Schroth MN (1975) Genetic transfer of Pseudomonas aeruginosa R factors to plant pathogenic Erwinia species. J Bacteriol 122:192–198

    Google Scholar 

  • Covey C, Richardson D, Carbon J (1976) A method for the deletion of restriction sites in bacterial plasmid deoxyribonucleic acid. Mol Gen Genet 145:155–158

    Google Scholar 

  • Datta N, Hedges RW (1972) Host range of R factors. J Gen Microbiol 70:453–460

    Google Scholar 

  • Figurski D, Helinski DR (1979) Replication of an origin containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652

    Google Scholar 

  • Figurski DH, Pohlman RF, Bechhofer DH, Prince AS, Kelton CA (1982) The broad host range plasmid RK2 encodes multiple kil genes potentially lethal to Escherichia coli host cells. Proc Natl Acad Sci USA 79:1935–1939

    Google Scholar 

  • Ingram LC, Richmond MH, Sykes RB (1973) Molecular characterization of the R factors implicated in the carbenicillin resistance of a sequence of Pseudomonas aeruginosa strains isolated from burns. Antimicrob Agents Chemother 3:279–288

    Google Scholar 

  • Kahn M, Kolter R, Thomas C, Figurski D, Meyer R, Remaut E, Helinski DR (1979) Plasmid cloning vehicles derived from plasmids ColE1, F, R6K and RK2. Methods Enzymol 68:268–280

    Google Scholar 

  • Kolter R, Helinski DR (1978) construction of plasmid R6K derivatives in vitro: characterization of the R6K replication region. Plasmid 1:571–580

    Google Scholar 

  • Komai N, Nishizawa T, Hayakawa Y, Murotsu T, Matsubara K (1982) Detection and mapping of six miniF-encoded proteins by cloning analysis of dissected miniF segments. Mol Gen Genet 186:193–203

    Google Scholar 

  • McKenny K, Shimatke H, Court D, Schmeissner U, Brady C, Rosenberg M (1981) A system to study promoter and terminator signals recognised by Escherichia coli RNA polymerase. In: Chirikjian JC, Papas TS (eds) Gene amplification and analysis, vol II. Analysis of nucleic acids by enzymatic methods. Elsevier/North Holland, Amsterdam, p 383–415

    Google Scholar 

  • Meyer R, Figurski D, Helinski D (1977) Physical and genetic studies with restriction endonucleases on the broad host-range plasmid RK2. Mol Gen Genet 152:129–135

    Google Scholar 

  • Meyer RJ (1979) Expression of incompatibility by derivatives of the broad host range IncP-1 plasmid RK2. Mol Gen Genet 177:155–161

    Google Scholar 

  • Olsen RH, Shipley P (1973) Host range and properties of the Pseudomons aeruginosa R factor R1822. J Bacteriol 113:772–780

    Google Scholar 

  • Sakanyan VA, Yakubor LZ, Alikhanian SI, Stepanov AI (1978) Mapping of RP4 plasmid using deletion mutants of pAS8 hybrid (RP4-ColE1). Mol Gen Genet 165:331–341

    Google Scholar 

  • Schmidhauser TJ, Filutowicz M, Helinski DR (1983) Replication of derivatives of the broad host-range plasmid RK2 in two distantly related bacteria. Plasmid 9:No 3

  • Selveraj G, Iyer VN (1981) Genetic transformation of Rhizobium meliloti by plasmid DNA. Gene 15:279–283

    Google Scholar 

  • Sutcliffe JG (1978) pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long. Nucl Acids Res 5:2721–2728

    Google Scholar 

  • Thomas CM, Meyer R, Helinski DR (1980) Regions of broad host-range plasmid RK2 which are essential for replication and maintenance. J Bacteriol 141:213–222

    Google Scholar 

  • Thomas CM (1981a) Molecular genetics of broad host range plasmid RK2. Plasmid 5:10–19

    Google Scholar 

  • Thomas CM (1981b) Complementation analysis of replication and maintenance functions of broad host range plasmids RK2 and RP1. Plasmid 5:277–291

    Google Scholar 

  • Thomas CM, Hussain AAK, Smith CA (1982) Maintenance of broad host range plasmid RK2 replicons in Pseudomonas aeruginosa. Nature 298:674–676

    Google Scholar 

  • Tolun A, Helinski DR (1982) Separation of the minimal replication region of the F plasmid into a replication origin segment and a trans-acting segment. Mol Gen Genet 186:372–377

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Genetics, University of Birmingham, P.O. Box 363, B15 2TT, Birmingham, England

    Christopher A. Smith & Christopher M. Thomas

Authors
  1. Christopher A. Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christopher M. Thomas
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by D. Sherratt

Rights and permissions

Reprints and permissions

About this article

Cite this article

Smith, C.A., Thomas, C.M. Deletion mapping of kil and kor functions in the trfA and trfB regions of broad host range plasmid RK2. Mol Gen Genet 190, 245–254 (1983). https://doi.org/10.1007/BF00330647

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330647

Keywords

Search

Navigation