Summary
A segment of DNA encoding incompatibility on the inc P-1 plasmid pRK248 was identified by the analysis of deletions generated in vitro, and then cloned into several unrelated and mutually compatible plasmids. These derivatives were tested for expression of P-1 incompatibility. It was demonstrated by transformation experiments that P-1 plasmids were efficiently eliminated from an E. coli host following introduction of any one of the derivatives. However, all the derivatives were compatible with each other. The cloned segment of pRK248 DNA is itself capable of autonomous replication, without being cloned into any plasmid, if plasmid-specified gene products are provided in trans. This satellite plasmid is eliminated from the cell by the inc P-1 plasmid pRK286. The results argue against a partitioning mechanism as the basis for P-1 incompatibility but are consistent with incompatibility being the consequence of negative regulation of copy number. For the inc P-1 system, susceptibility of the plasmid to elimination, but not its ability to eliminate, requires that the P-1 replication system is active.
Similar content being viewed by others
References
Allet, B., Bukhari, A.: Analysis of bacteriophage Mu and λ-Mu hybrid DNAs by specific endonucleases. J. Mol. Biol. 92, 529–540 (1975)
Armstrong, K., Helinski, D.R.: Restriction map of the ColE1 derivative pCR1. In: DNA Insertion elements, plasmids and episomes (Bukhari, A.I., Shapiro, J.A., Adhya, S.L., eds.), p. 681. New York: Cold Spring Harbor Laboratory 1977
Bachmann, B.: Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol. Rev. 36, 525–557 (1972)
Beringer, J.G.: R factor transfer in Rhizobium leguminosarum. J. Gen. Microbiol. 84, 188–198 (1974)
Cabello, F., Timmis, K., Cohen, S.N.: Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature 259, 285–290 (1976)
Cohen, S.N., Chang, A.C.Y., Hsu, L.: Non-chromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc. Natl. Acad. Sci. U.S.A. 69, 2110–2114 (1972)
Cohen, S.N., Chang, A.C.Y., Boyer, H.W., Helling, R.B.: Construction of biologically functional bacterial plasmids in vitro. Proc. Natl. Acad. Sci. U.S.A. 70, 3240–3244 (1973)
Covey, C., Richardson, D., Carbon, J.: A method for the deletion of restriction sites in bacterial plasmid deoxyribonucleic acid. Mol. Gen. Genet. 145, 155–158 (1976)
DeVries, J., Maas, W.: Description of an incompatibility mutant of Escherichia coli. J. Bacteriol. 115, 312–220 (1973)
Figurski, D., Kolter, R., Meyer, R., Kahn, M., Eichenlaub, R., Helinski, D.R.: Replication regions of plasmids ColE1, F, R6K and RK2. In: Microbiology 1978 (Schlesinger, R., ed.), pp. 105–109. Washington American Society for Microbiology 1978
Hedges, R.W., Jacob, A.E.: Transposition of ampicillin resistance from RP4 to other replicons. Mol. Gen. Genet. 132, 31–40 (1974)
Heffron, F., Bedinger, P., Champoux, J., Falkow, S.: Deletions affecting transposition of an antibiotic resistance gene. In: DNA insertion elements, plasmids and episomes (Bukhari, A.I., Shapiro, J.A., Adhya, S.L., eds.) pp. 161–167. New York: Cold Spring Harbor Laboratory 1977
Ingram, L.C., Richmond, M.H., Sykes, R.B.: Molecular characterization of the R factors implicated in the carbenicillin resistance of a sequence of Pseudomonas aeruginosa strains isolated form burns. Antimicrob. Agents Chemother. 3, 279–288 (1973)
Katz, L., Kingsbury, D.T., Helinski, D.R.: Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-Protein relaxation complex. J. Bacteriol. 114, 577–591 (1973)
Kleckner, N.: Translocatable elements in procaryotes. Cell 11, 11–23 (1977)
Kolter, R., Helinski, D.R.: Activity of the replication terminus of plasmid R6K in hybrid replicons in Escherichia coli. J. Mol. Biol. 124, 425–441 (1978)
Lovett, M.A., Helinski, D.R.: Method for the isolation of the replication region of a bacterial replicon: construction of a mini F′Km plasmid. J. Bacteriol. 127 982–987 (1976)
Manis, J., Kline, B.: F incompatibility and copy number genes: their map locations and interactions. Plasmid 1, 492–507 (1978)
Meyer, R., Figurski, D., Helinski, D.R.: Molecular vehicle properties of the broad host range plasmid RK2. Science 190, 1226–1228 (1975)
Meyer, R., Figurski, D., Helinski, D.R.: Physical and genetic studies with restriction endonucleases on the broad host-range plasmid RK2. Mol. Gen. Genet. 152, 129–135 (1977a)
Novick, R.P., Schwesinger, M.: Independence of plasmid incompatibility and replication control functions in Staphylococcus aureus. Nature 262, 623–626 (1976)
Palchaudhuri, S., Maas, W.K.: Physical mapping of a DNA sequence common to plasmids of incompatibility group F-1. Proc. Natl. Acad. Sci. U.S.A. 74, 1190–1194 (1977)
Panopoulos, N.J., Cho, J.J., Guimares, W.V., Schroth, M.N.: Genetic transfer of Pseudomonas aeruginosa R factors to Pseudomonas and Erwinia plant pathogens. In: Proceedings of the first intersectional congress of IAMS (Hasegawa, T., ed.), Vol 1, pp. 142–161. Tokyo: Science Council of Japan 1975
Pritchard, R.H.: Control of DNA replication in bacteria. In: DNA Synthesis present and future (Molineux, I., Kohiyama, M., eds.), pp. 1–26. New York, London: Plenum Press 1978
Roberts, R.J.: Appendix D: Restriction Endonucleases. In: DNA insertion elements, plasmids and episomes (Bukhari, A.I., Shapiro, J.A., Adhya, S.L., eds.), pp. 757–768. New York: Cold Spring Harbor Laboratory 1977
Rownd, R.: Replication of a bacterial episome under relaxed control. J. Mol. Biol. 44, 387–402 (1969)
Shapiro, J.A., Sporn, P.: Tn402: a new transposable element determining trimethoprim resistance that inserts in bacteriophage lambda. J. Bacteriol. 129, 1632–1635 (1977)
Timmis, K., Andres, I., Slocombe, P.M.: Plasmid incompatibility: cloning analysis of an inc FII determinant of R6-5. Nature 273, 27–32 (1978)
Uhlin, B.E., Nordstrom, K.: R plasmid gene dosage effects in Escherichia coli K-12: copy mutants of the R plasmid R1 drd-19. Plasmid 1, 1–7 (1977)
Author information
Authors and Affiliations
Additional information
Communicated by H.W. Boyer
Rights and permissions
About this article
Cite this article
Meyer, R.J. Expression of incompatibility by derivatives of the broad host-range inc P-1 plasmid RK2. Molec. Gen. Genet. 177, 155–161 (1979). https://doi.org/10.1007/BF00267265
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00267265