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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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Summary

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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.

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

References

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