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The phasmid as a tool for plasmid genetics: II. Isolation of point mutations that affect replication of a ColE1-related plasmid

Published online by Cambridge University Press:  14 April 2009

Gianni Cesareni ,
Luisa Castagnoli  and
Sydney Brenner
Gianni Cesareni
Affiliation:
MRC Laboratory of Molecular Biology, University Postgraduate Medical School, Hills Road, Cambridge CB2 2QH, England
Luisa Castagnoli
Affiliation:
MRC Laboratory of Molecular Biology, University Postgraduate Medical School, Hills Road, Cambridge CB2 2QH, England
Sydney Brenner
Affiliation:
MRC Laboratory of Molecular Biology, University Postgraduate Medical School, Hills Road, Cambridge CB2 2QH, England
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The insertion of a high-copy-number plasmid into a lambdoid phage chromosome which lacks a functional repressor gene confers on the hybrid ‘phasmid’ the capacity to grow on an immune lysogen. This was found to be due to titration of repressor because of plasmid replication. We have exploited this property in order to isolate mutants that affect plasmid replication. These mutants have been mapped in a region that was previously characterized as necessary for plasmid replication and incompatibility properties. Some of the mutations could revert at frequencies characteristic of single-base-pair change mutations.

Information

Type
Research Article
Information
Genetics Research , Volume 40 , Issue 3 , December 1982 , pp. 233 - 247
Copyright
Copyright © Cambridge University Press 1982

References

REFERENCES

Backman, K., Betlach, M., Boyer, H. W. & Yanofsky, S. (1978). Genetic and physical studies on the replication of ColE1-type plasmids. Cold Spring Harbor Symposia on Quantitative Biology 43, 69–76.CrossRefGoogle Scholar
Brenner, S., Cesareni, G. & Karn, J. (1982). Phasmids: hybrids between ColE1 plasmids and bacteriophage lambda. Gene 17, 27–44.CrossRefGoogle ScholarPubMed
Castagnoli, L., Cesareni, G. & Brenner, S. (1982). The phasmid as a tool for plasmid genetics. I. Fine structure of the β-lactamase gene. Genetical Research 40, 217–231.CrossRefGoogle Scholar
Clewell, D. B. (1972). Nature of ColE1 plasmid replication in Escherichia coli in the presence of chloramphenicol. Journal of Bacteriology 139, 667–676.CrossRefGoogle Scholar
Donoghue, D. J. & Sharp, P. A. (1978). Replication of ColE1 plasmid DNA in vivo requires no plasmid encoded proteins. Journal of Bacteriology 133, 1287–1294.CrossRefGoogle Scholar
Hashimoto-Gotoh, T. & Inselburg, J. (1979). Isolation and characterization of replication deficient mutants of ColE1 plasmids. Journal of Bacteriology 139, 597–607.CrossRefGoogle ScholarPubMed
Herschfield, V., Boyer, H. W., Chow, L. & Helinski, D. R. (1976). Characterization of a mini-ColE1 plasmid. Journal of Bacteriology 126, 447–453.CrossRefGoogle Scholar
Itoh, T. & Tomizawa, J. (1978). Initiation of replication of plasmid ColE1 DNA by RNA polymerase, Ribonuclease H, and DNA polymerase I. Cold Spring Harbor Symposia on Quantitative Biology 43, 409–418.CrossRefGoogle Scholar
Kahn, M. & Helinski, D. (1978). Construction of a novel plasmid-phage hybrid: use of the hybrid to demonstrate ColE1 DNA replication in vivo in the absence of a ColE1 specified protein. Proceedings of the National Academy of Sciences, U.S.A. 75, 2200–2204.CrossRefGoogle ScholarPubMed
Muesing, M., Tam, J., Shepard, H. M. & Polinsky, B. (1981). A single base pair alteration is responsible for the DNA overproduction phenotype of plasmid copy number mutant. Cell 24, 235–242.CrossRefGoogle ScholarPubMed
Mukai, T., Ohkubo, H., Shimada, K. & Takagi, Y. (1978). Isolation and characterization of a plaque forming lambda bacteriophage carrying a ColE1 plasmid. Journal of Bacteriology 135, 171–177.CrossRefGoogle ScholarPubMed
Naito, S. & Uchida, H. (1980). Initiation of DNA replication in a ColE1 type plasmid: isolation of mutations in the ori region. Proceedings of the National Academy of Sciences of the USA 77, 6744–6748.CrossRefGoogle Scholar
Ohmori, H. & Tomizawa, J. (1979). Nucleotide sequence in the region for maintenance of colicin E1 plasmid. Molecular and General Genetics 176, 161–170.CrossRefGoogle ScholarPubMed
Oka, A., Nomura, M., Sugisaki, H., Sugimoto, K. & Takanami, M. (1979). Nucleotide sequence of small ColE1 derivatives: structure of the regions essential for autonomous replication and colicin E1 immunity. Molecular and General Genetics 172, 151–159.CrossRefGoogle ScholarPubMed
Reichardt, L. F. (1975). Control of bacteriophage lambda represser synthesis: regulation of the maintenance pathway by the cro and cl products. Journal of Molecular Biology 93, 289–309.CrossRefGoogle Scholar
Shafferman, A., Cohen, S. & Flashner, Y. (1978). A DNA segment within colicin E1 structural gene on ColE1 affecting immunity to colicin. Molecular and General Genetics 164, 259–264.CrossRefGoogle ScholarPubMed
Shepard, H. M., Gelfand, D. M. & Polinski, B. (1979). Analysis of a recessive plasmid copy number mutant: evidence for negative control of ColE1 replication. Cell 18, 267–275.CrossRefGoogle Scholar
Tomizawa, J., Sakakibara, Y. & Kakefuda, T. (1975). Replication of colicin E1 plasmid DNA added to cell extracts. Proceedings of the National Academy of Sciences of the USA 72, 1050–1054.CrossRefGoogle ScholarPubMed
Tomizawa, J., Itoh, T., Selzer, G. & Sam, T. (1981). Inhibition of ColE1 primer formation by a plasmid specified small RNA. Proceedings of the National Academy of Sciences of the USA 78, 1421–1425.CrossRefGoogle ScholarPubMed
Uhlin, B. E. & Nördstrom, K. (1977). R plasmid gene dosage effects in Escherichia coli K12: copy mutants of the R plasmid Rldrd19. Plasmid 1, 1–7.CrossRefGoogle Scholar
Windass, J. D. & Brammar, W. J. (1979). Aberrant immunity behaviour of hybrid imm21 phages containing the DNA of ColE1-type plasmids. Molecular and General Genetics 172, 329–337.CrossRefGoogle ScholarPubMed