Summary
A 370 base pair (bp) fragment of R1162 DNA encoding the incompatibility determinant has been cloned and sequenced. The DNA is located between 6.1 and 6.5 on the R1162 map, near the origin of replication. The sequence contains three perfectly conserved 20 bp direct repeats, with 11 bp of this sequence repeated a fourth time. The direct repeat unit shows some homology with that of another, unrelated broad host-range plasmid, RK2. The cloned DNA has two other properties: it lowers the copy number of R1162 when cloned into this plasmid, and it is required in cis for replication of R1162 satellite plasmids.
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References
Barth PT, Grinter NJ (1974) Comparison of the deoxyribonucleic acid molecular weights and homologies of plasmids conferring linked resistance to streptomycin and sulfonamides. J Bacteriol 120:618–630
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7:1512–1523
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles. II. A multiple cloning system. Gene 2:95–113
Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the p15A cryptic miniplasmid. J Bacteriol 134:1141–1156
Churchward G, Linder P, Caro L (1983) The nucleotide sequence of replication and maintenance functions encoded by plasmid pSC101. Nucl Acids Res 11:5645–5659
Cohen SN, Chang ACY, Hsu L (1972) Non-chromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R factor DNA. Proc Natl Acad Sci USA 69:2100–2104
Germino J, Bastia D (1983) Interaction of plasmid R6K-encoded replication initiator protein with its binding sites on DNA. Cell 34:125–134
Hershfield V, Boyer HW, Yanofsky C, Lovett MA, Helinski DR (1974) Plasmid ColE1 as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci USA 71:3455–3459
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197
Jobanputra RS, Datta N (1974) Trimethoprim R factors in enterobacteria from clinical specimens. J Med Microbiol 7:169–177
Kahn M, Helinski DR (1978) Construction of a novel plasmidphage hybrid: use of the hybrid to demonstrate ColE1 DNA replication in vivo in the absence of a ColE1-specified protein. Proc Natl Acad Sci USA 75:2200–2204
Kamio Y, Terawaki Y (1983) Nucleotide sequence of an incompatibility region of mini-Rts1 that contains five direct repeats. J Bacteriol 155:1185–1191
Kolter R (1981) Replication properties of plasmid R6K. Plasmid 5:2–9
Kolter R, Helinski DR (1982) Plasmid R6K DNA replication II. Direct nucleotide sequence repeats are required for an active ψ-origin. J Mol Biol 161:45–56
Lovett M, 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
Marko MA, Chipperfield R, Birnboim HC (1982) A procedure for the large-scale isolation of highly purified plasmid DNA using alkaline extraction and binding to glass powder. Anal Biochem 121:382–387
Maxam AM, Gilbert W (1980) Sequencing end-labelled DNA with base-specific chemical cleavages. In: Grossman L, Moldave K (eds) Methods in enzymology, vol 75. Academic Press, New York, pp 499–560
Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene 19:269–276
Messing J, Crea R, Seeburg PH (1981) A system for shotgun DNA sequencing. Nucl Acids Res 9:309–321
Meyer R, Shapiro J (1980) Genetic organization of the broad host-range IncP-1 plasmid R751. J Bacteriol 143:1362–1373
Meyer R, Hinds M, Brasch M (1982) Properties of R1162 a broadhost range, high-copy-number plasmid. J Bacteriol 150: 552–562
Murotsu T, Matsubara K, Sugisaki H, Takanami T (1981) Nine unique repeating sequences in a region essential for replication and incompatibility of the mini-F plasmid. Gene 15:257–271
Pritchard RH (1978) Control of DNA replication in Bacteria. In: Molineux I, Kohiyama M (eds) DNA synthesis present and future. Plenum Press, New York, London, pp 1–26
Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353
Sanger F, Nicklen S, Coulson R (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Shafferman A, Kolter R, Stalker D, Helinski DR (1982) Plasmid R6K DNA replication. III. Regulatory properties of the initiation protein. J Mol Biol 161:57–76
Shine J, Dalgarno K (1974) The 3′ terminal sequence of E. coli 16S rRNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346
Stalker DM, Kolter R, Helinski DR (1979) Nucleotide sequence of the region of the origin of replication of the antibiotic resistance plasmid R6K. Proc Natl Acad Sci USA 76:1150–1154
Stalker DM, Thomas CM, Helinski DR (1981) Nucleotide sequence of the region of the origin of replication of the broad host-range plasmid RK2. Mol Gen Genet 181:8–12
Stougaard P, Molin S, Nordstrom K (1981) RNAs involved in copy-number control and incompatibility of plasmid R1. Proc Natl Acad Sci USA 78:6008–6012
Thomas CM, Stalker DM, Helinski DR (1981) Replication and incompatibility properties of segments of the origin region of replication of the broad host-range plasmid RK2. Mol Gen Genet 181:1–7
Tolun A, Helinski DR (1981) Direct repeats of the F plasmid incC region express F incompatibility. Cell 24:687–694
Tomizawa J-I, Itoh T, Selzer G, Som T (1981) Inhibition of ColE1 RNA primer formation by a plasmid-specified small RNA. Proc Natl Acad Sci USA 78:1421–1425
Tsutsui H, Fujiyama A, Murotsu T, Matsubara K (1983) Role of nine repeating sequences of the mini-F genome for expression of F-specific incompatibility phenotype and copy number control. J Bacteriol 155:337–344
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Communicated by D. Sherratt
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Lin, Ls., Meyer, R.J. Nucleotide sequence and functional properties of DNA encoding incompatibility in the broad host-range plasmid R1162. Molec. Gen. Genet. 194, 423–431 (1984). https://doi.org/10.1007/BF00425554
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DOI: https://doi.org/10.1007/BF00425554