Summary
Specialized transducing lambda phages, λoriR1, harboring DNA from the resistance plasmid R1drd-19 and its copy mutant pKN103 were isolated. From measurements of CCC-DNA content it is concluded that upon infection the phages can establish themselves as self-replicating plasmids in recA hosts lysogenic for lambda. It is thought that this bypassing of lambda immunity is due to the presence of the R1 origin of replication. The plasmids are sensitive to the incompatibility expressed by plasmid R1. This has been shown mainly by transduction of λoriR1 into recipients containing R1 plasmids or plasmid pBR322 carrying the basic replicon. We were able to demonstrate that a copy mutant of plasmid R1 was insensitive to copA +, but sensitive to the conserted action of Pst1 fragments F1 and F2. This mutant was previously assumed to be of the dominant type. Physical mapping of the λoriR1 derivatives verified that they carry the basic replicon of plasmid R1. The plasmids are not stably maintained, but are lost in a frequency of 1%–2% per cell generation, which is consistent with their lack of the R1par region.
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References
Bachmann BJ, Low BK (1980) Linkage map of E. coli K-12. Microbiol Rev 44:1–56
Beard JP, Conolly JC (1975) Detection of a protein, similar to the sex pilus subunit, in the outer membrane of Escherichia coli cells carrying a derepressed F-like R-factor. J Bacteriol 122:59–65
Bertani G (1951) Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol 62:293–300
Blohm D, Goebel W (1978) Restriction map of the antibiotic resistance plasmid R1drd-19 and its derivatives pKN102 (R1drd-19B2) and R1drd-16 for the enzymes BamH1, HindIII, EcoR1 and SalI. Mol Gen Genet 167:119–127
Boman HG, Eriksson-Grennberg KG, Normark S, Matsson E (1968) Resistance of Escherichia coli to penicillins, IV. Genetic study of mutants resistant to DL-ampicillin concentrations of 100 μ/ml. Genet Res 12:169–185
Clark JD, Maaløe O (1967) DNA replication and the division cycle in Escherichia coli. J Mol Biol 23:99–112
Clewell DB, Helinski DR (1969) Supercoiled circular DNA-protein complex in E. coli: Purification and induced conversion to an open circular DNA form. Proc Natl Acad Sci USA 62:1159–1166
Couturier M, Janssens J, Bex F, Desmyter A, Bonnevalle I (1979) Construction in vitro of “phage-plasmid” chimerae: A new tool to analyse the mechanism of plasmid maintenance. Mol Gen Genet 169:113–116
Dempsey WB, Willetts NS (1975) Plasmid co-integrates of prophage lambda and R factor R100. J Bacteriol 26:166–176
Dempsey WB, McIntire SA (1979) Lambda transducing phages derived from a finO− R100::λ cointegrate plasmid: Proteins encoded by the R100 replication/incompatibility region and the antibiotic resistance determinant. Mol Gen Genet 176:319–334
Enquist LW, Skalka A (1973) Replication of bacteriophage λDNA dependent on the function of host and viral genes. J Mol Biol 75:185–212
Hayakawa Y, Matsubara K (1979) Construction and some properties of packageable plasmid F. Mol Gen Genet 169:107–112
Kollek R, Oertel W, Goebel W (1978) Isolation and characterization of the minimal fragment required for autonomous replication of a copy mutant (pKN102) of the resistance factor R1. Mol Gen Genet 162:51–58
Matsubara K (1976) Genetic structure and regulation of a replicon of plasmid λdv. J Mol Biol 102:427–439
Meyenburg Kv, Hansen FG, Nielsen LD, Riise E (1978) Origin of replication, oriC, of the E. coli chromosome on specialized transducing phages λasn. Mol Gen Genet 160:287–295
Meynell E, Datta N (1967) Mutant drug-resistant factors of high transmissibility. Nature 214:885–887
Miller J (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Molin S, Stougaard P, Uhlin BE, Gustafsson P, Nordström K (1979) Clustering of genes involved in replication, copy number control, incompatibility and stable maintenance of the resistance plasmid R1drd-19. J Bacteriol 138:70–79
Molin S, Stougaard P, Nordström K (1980) Control of replication of plasmid R1. In: Schlessinger D (ed) Microbiology-1980 Washington DC: Am Soc Microbiol (in press)
Molin S, Stougaard P, Light J, Nordström M, Nordström K (1981) Isolation and characterization of new copy mutants of plasmid R1, and identification of a polypeptide incolved in copy number control. Mol Gen Genet 181:123–130
Molin S, Nordström K (1980) Control of replication of plasmid R1. II. Functions involved in replication, copy number control, incompatibility and switch-off of replication. J Bacteriol 141:111–120
Mukai T, Ohkubo H, Shimada K, Takagi Y (1978) Isolation and characterization of a plaque-forming lambda bacteriophage carrying a ColEI plasmid. J Bacteriol 135:171–177
Nordström K, Eriksson-Grennberg KG, Boman HG (1968) Resistance of E. coli to penicillins III. AmpB, a locus affecting episomally and chromosomally mediated resistance to ampicillin and chloramphenicol. Genet Res 12:157–168
Nordström K, Molin S, Aagaard-Hansen H (1980) Partitioning of plasmid R1 in Escherichia coli. I. Kinetics of loss of plasmid derivatives deleted of the par region. Plasmid 3:218–227
Novick RP, Clowes RC, Cohen SN, Curtiss R III, Datta N, Falkow S (1976) Uniform nomenclature for bacterial plasmids: A proposal. Bacteriol Rev 40:168–189
Rosen J, Ryder T, Inokuchi H, Ohtsubo H, Ohtsubo E (1980) Genes and sites involved in replication and incompatibility of an R100 plasmid derivative based on nucleotide sequence analysis. Mol Gen Genet 179:527–537
Schrenk K, Weisberg RA (1975) A simple method for making new transducing lines of coliphage λ. Mol Gen Genet 137:101–107
Stougaard P, Molin S, Nordström K, Hansen FG (1980) The nucleotide sequence of the replication control region of the resistance plasmid R1drd-19. Mol Gen Genet 181:116–122
Synenki RM, Nordheim A, Timmis KN (1979) Plasmid replication functions. III. Origin and direction of replication of a “Mini” plasmid derived from R6-5. Mol Gen Genet 168:27–36
Uhlin BE, Nordström K (1975) Plasmid incompatibility and control of replication: Copy mutants of the R-factor R1 in Escherichia coli K-12. J Bacteriol 124:641–649
Uhlin BE, Nordström K (1977) R plasmid gene dosage effects in Escherichia coli K-12: Copy mutants of the R plasmid R1drd-19. Plasmid 1:1–7
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Communicated by D. Sherratt
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Dreisig, H., Riise, E. & Nordström, K. Isolation and characterization of lambda phages carrying the basic replicon of the resistance plasmid R1. Molec. Gen. Genet. 182, 148–153 (1981). https://doi.org/10.1007/BF00422782
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DOI: https://doi.org/10.1007/BF00422782