Summary
The mini plasmids deriving from pKN102, a copy mutant of the antibiotic resistance factor R1drd-19 of E. coli, share a common DNA sequence of 2.6 kb, which carries the minimal functions for autonomous replication. By cloning of two PstI fragments of this region it could be demonstrated that the “basic replicon” is a DNA segment not larger than 1.8 kb, which carries the origin of replication and the genetic information for at least two proteins. Protein F (MW=11.000 dalton) seems to be synthesed in larger amounts in minicells of E. coli than protein C (20.000 dalton). Plasmids containing this isolated replicon of R1 are completely compatible with the parental plasmid R1drd-19.
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Boidol, W., Siewert, G., Lindenmaier, W., Luibrand, G., Goebel, W.: Properties of hybrid plasmids, consisting of parts of the mini-R1 factor Rsc11 and ColE1. Molec. gen. Genet. 152 231–237 (1977)
Bonner, W.M., Laskey, R.A.: A film detection method of tritium-labeled proteins and nucleic acids in polyacrylamide gels. Europ. J. Biochem. 46, 83–88 (1974)
Cohen, S.N., Chang, A.C.Y., Boyer, H.W., Helling, R.B.: Construction of biologically functional bacterial plasmids in vitro. Proc. nat. Acad. Sci. (Wash.) 70, 3240–3244 (1973)
Cohen, S.N.: Transposable genetic elements and plasmid evolution. Nature (Lond.) 72, 3628–3632 (1975)
Goebel, W., Bonewald, R.: Class of small multicopy plasmids originating from the mutant antibiotic resistance factor R1 drd-19B2. J. Bact. 123, 658–665 (1975)
Goebel, W., Lindenmaier, W., Schrempf, H., Kollek, R., Blohm, D.: Dissociation and Recombination of fragments with defined functions of the antibiotic resistance factor R1. Sandoz-Symp. “R-Factors” — Their Properties and Possible Control, Vienna, 1977, pp. 261–273
Goebel, W., Lindenmaier, W., Pfeifer, F., Schrempf, H., Schelle, B.: Transposition and insertion of intact deleted and enlarged ampicillin transposon Tn3 from mini R1 (Rsc) plasmids into transfer factors. Molec. gen. Genet. 157, 119–129 (1977)
Levy, S.B.: R-factor proteins synthesized in E. coli minicells: incorporation studies with different R factors and detection of deoxyribonucleicacid binding proteins. J. Bact. 120, 1451–1463 (1974)
Luibrand, G., Blohm, D., Mayer, H., Goebel, W.: Characterization of small ampicillin resistance plasmids (Rsc) originating from the mutant antibiotic resistance factor R1drd-19B2. Molec. gen. Genet. 152, 43–51 (1977)
Mayer, H., Luibrand, G., Goebel, W.: Replication of the mini-R1 plasmid Rsc11 and Rsc11 hybrid plasmids. Molec. gen. Genet. 152, 145–152 (1977)
Nordström, K., Ingram, L.C., Lundbäck, K.: Mutations in R-factors of escherichia coli causing an increased number of R-factor copies per chromosome. J. Bact. 110, 562–569 (1974)
Ohtsubo, E., Rosenbloom, M., Schrempf, H., Goebel, W., Rosen, J.: Site specific recombination involved in the generation of small plasmids. Molec. gen. Genet. 159, 131–141 (1978)
Perlman, D., Rownd, R.H.: Two origins of replication in composite R plasmid DNA. Nature (Lond.) 259, 281–284 (1976)
Pritchard, R.H., Barth, P.T., Collins, J.: Control of DNA synthesis in bacteria. Symp. Soc. gen. Microbiol. 19, 263–297 (1969)
Sharp, P.A., Cohen, S.N., Davidson, N.: Electron microscope heteroduplex studies of sequence relations among plasmids of E. coli. II Structure of drug resistance (R) factors and F factors. J. molec. Biol. 75, 235–255 (1973)
Silver, L., Chandler, M., Caro, L.: Origin and direction of replication of the drug resistance plasmid R100.1 and of an RTF derivative in synchronized cultures. J. Bact. (in press)
Timmis, K., Cabello, F., Cohen, S.N.: Cloning, isolation and characterization of replication regions of complex plasmid genomes. Proc. nat. Acad. Sci. (Wash.) 72, 2242–2246 (1975)
Uhlin, B.E., Nordström, K.: Plasmid incompatibility and control of replication: Copy mutants of the R-factor R1 in Escherichia coli K12. J. Bact. 124, 641–649 (1975)
Uhlin, B.E., Gustafson, P., Molin, S., Nordström, K.: Copy number control mutants of the R plasmid R1 in Escherichia coli. DNA synthesis: present and future (M. Kohiyama and I. Molineux, ed.), New York: Plenum Publishing Corp. (in press)
Yoshikawa, M.: Identification and mapping of the replication genes of an R factor, R100.1, integrated into the chromosome of Escherichia coli K12. J. Bact. 118, 1123–1131 (1974)
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Communicated by F. Kaudewitz
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Kollek, R., Oertel, W. & Goebel, W. Isolation and characterization of the minimal fragment required for autonomous replication (“Basic replicon”) of a copy mutant (pKN102) of the antibiotic resistance factor R1. Molec. Gen. Genet. 162, 51–57 (1978). https://doi.org/10.1007/BF00333850
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DOI: https://doi.org/10.1007/BF00333850