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Plasmid mini-F encoded proteins

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Summary

Proteins specified by the mini-F plasmid (EcoRI restriction fragment f5) were labeled in Escherichia coli minicells and analyzed by SDS-PAGE. Four mini-F encoded proteins could be identified, having molecular weights of 44,000 (A), 36,000 (B), 34,000 (C), and 25,300 (D) daltons. The absence of certain proteins in deleted derivatives of mini-F, generated by treatment with various restriction endonucleases, allowed mapping of the proteins. The A protein maps between F-coordinates 45.7 and 47.9 kb. The gene locus for the B protein is located between 47.2 and 49.3 kb. The C protein maps on a BamHI fragment bordered by F-coordinates 41.5 and 42.8 kb, and finally the D protein maps between 42.8 and 43.8 kb. In addition our data confirm that there are two incompatibility loci on the mini-F genome, located between 45.7 and 47.2 kb (incA) and 44.0 and 45.7 kb (incB).

We suggest that (i) the C and D proteins are positive control elements, interacting with origin I and origin II, respectively, (ii) that the incB locus is involved in plasmid partitioning, and (iii) that the A protein encoded by the incA locus is a negative control element.

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References

  • Adler HI, Fisher WD, Cohen A, Hardigree AA (1966) Miniature Escherichia coli cells deficient in DNA. Proc Natl Acad Sci USA 57:321–326

    Google Scholar 

  • Andrés I, Slocombe PM, Cabello F, Timmis JK, Lurz R, Burkhardt HJ, Timmis KN (1979) Plasmid replication functions. II. Cloning analysis of the RepA replication region of antibiotic resistance plasmid R6-5 Mol Gen Genet 168:1–25

    Google Scholar 

  • Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36:525–557

    Google Scholar 

  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113

    Google Scholar 

  • Bonner WM, Laskey RA (1974) A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 46:83–88

    Google Scholar 

  • Cabello F, Timmis K, Cohen SN (1976) Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature 259:285–290

    Google Scholar 

  • Dougan E, Sherratt D (1977) The transposon Tn1 as a probe for studying ColE1 structure and function. Mol Gen Genet 151:151–160

    Google Scholar 

  • Eichenlaub R (1979) Mutants of the mini-F plasmid pML31 thermosensitive in replication. J Bacteriol 138:559–566

    Google Scholar 

  • Eichenlaub R, Figurski D, Helinski DR (1977) Bidirectional replication from a unique origin in a mini-F plasmid. Proc Natl Acad Sci USA 74:1138–1141

    Google Scholar 

  • Figurski D, Kolter R, Meyer R, Kahn M, Eichenlaub R, Helinski DR (1978) Replication regions of plasmids ColE1, F, R6K, and RK2. In: Schlessinger D (ed) Microbiology-1978. Am Soc Micro, Washington D.C., pp 105–109

    Google Scholar 

  • Gustafsson P, Nordström K (1978) Temperature-dependent and amber copy mutants of plasmid Rldrd-19 in Escherichia coli. Plasmid 1:134–144

    Google Scholar 

  • Guyer MS, Figurski D, Davidson N (1976) Electron microscope study of a plasmid chimera containing the replication region of the Escherichia coli F plasmid. J Bacteriol 127:988–997

    Google Scholar 

  • Inuzuka M, Helinski DR (1978) Requirement of a plasmid encoded protein for the replication in vitro of plasmid R6K. Proc Natl Acad Sci USA 75:5381–5385

    Google Scholar 

  • Jacob F, Brenner S, Cuzin F (1963) On the regulation of DNA replication in bacteria. Cold Spring Harbor Symp Quant Biol 28:329–343

    Google Scholar 

  • Kahn ML, Figurski D, Ito L, Helinski DR (1978) Essential regions for replication of a stringent and a relaxed plasmid in Escherichia coli. Cold Spring Harbor Symp Quant Biol 43:99–103

    Google Scholar 

  • Kennedy N, Bentin L, Achtman M, Skurray R, Rahmsdorf U, Herrlich P (1977) Conjugation proteins encoded by the F sex factor. Nature 270:580–585

    Google Scholar 

  • Kingsbury DT, Helinski DR (1970) DNA polymerase as a requirement for the maintenance of the bacterial plasmid colicinogenic factor E1. Biochem Biophys Res Commun 41:1538–1544

    Google Scholar 

  • Kingsbury DT, Helinski DR (1973) Temperature-sensitive mutants for the replication of plasmids in Escherichia coli: Requirement for DNA polymerase I in the replication of the plasmid ColE1. J Bacteriol 114:1116–1124

    Google Scholar 

  • Kline BC (1979) Incompatibility between F'lac, R386, and F:pSC101 recombinant plasmids: The specificity of F incompatibility genes. Plasmid 2:437–445

    Google Scholar 

  • Kollek R, Oertel W, Goebel W (1978) Isolation and characterization of the minimal fragment required for autonomous replication (‘basic replicon’) of a copy mutant (pKN102) of the antibiotic resistance factor R1. Mol Gen Genet 162:51–57

    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 

  • Kolter R, Inuzuka M, Helinski DR (1978) Trans-complementation dependent replication of a low molecular weight origin fragment from plasmid R6K. Cell 15:1199–1208

    Google Scholar 

  • Laskey RA, Mills AD (1975) Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem 56:335–341

    Google Scholar 

  • Lovett MA, Helinski DR (1976) Method for the isolation of the replication region of a bacterial replicon: construction of a mini-F′ Km plasmid. J Bacteriol 127:982–987

    Google Scholar 

  • Lugtenberg B, Meijers J, Peters R, Hoek P, van der Alphen L (1975) Electrophoretic resolution of the “major outer membrane protein” of Escherichia coli K12 into four bands. FEBS Lett 58:254–258

    Google Scholar 

  • Manis JJ, Kline BC (1977) Restriction endonuclease mapping and mutagenesis of the F sex factor replication region. Mol Gen Genet 152:175–182

    Google Scholar 

  • Manis J, Kline BC (1978) F plasmid incompatibility and copy number genes: their map locations and interaction. Plasmid 1:492–507

    Google Scholar 

  • Novick RP, Hoppensteadt FC (1978) On plasmid incompatibility. Plasmid 1:421–434

    Google Scholar 

  • Palchaudhuri S, Maas WK (1977) Physical mapping of a DNA sequence common to plasmids of incompatibility group FI. Proc Natl Acad Sci USA 74:1190–1194

    Google Scholar 

  • Pritchard RH, Barth PT, Collins J (1969) Control of DNA synthesis in bacteria. Symp Soc Gen Microbiol 19:263–297

    Google Scholar 

  • Scherzinger F, Litfin F, Jost E (1973) Stimulation of T7 DNA polymerase by a new phage-coded protein. Mol Gen Genet 123:247–262

    Google Scholar 

  • Smith DI, Blattner FR, Davies J (1976) The isolation and partial characterization of a new restriction endonuclease from Providencia stuartii. Nucleic Acids Res 3:343–353

    Google Scholar 

  • Sutcliffe JG (1978) Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43:77–90

    Google Scholar 

  • Timmis KN, Cabello F, Cohen SN (1975) Cloning, isolation, and characterization of replication regions of complex plasmid genomes. Proc Natl Acad Sci USA 72:2242–2246

    Google Scholar 

  • Uhlin BE, Nordström K (1978) A runaway-replication mutant of plasmid R1drd-19: temperature-dependent loss of copy number control. Mol Gen Genet 165:167–179

    Google Scholar 

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Communicated by J. Schell

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Wehlmann, H., Eichenlaub, R. Plasmid mini-F encoded proteins. Molec. Gen. Genet. 180, 205–211 (1980). https://doi.org/10.1007/BF00267371

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