Amber-mutants of plasmid mini-F defective in replication |
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Authors: | Rudolf Eichenlaub and Hermann Wehlmann |
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Institution: | (1) Lehrstuhl Biologie der Mikroorganismen, Ruhr-Universität Bochum, Postfach 102148, D-4630 Bochum, Federal Republic of Germany |
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Abstract: | Summary Amber mutants of the mini-F plasmid pML31 were isolated with the mutagen hydroxylamine. Under non-permissive conditions amber mutants segregate and show no incorporation of label into supercoiled plasmid DNA in double-label experiments. Wild-type and one mutant of mini-F were integrated by recombinant DNA techniques into the single EcoRI site of plasmid vector pBR322. Plasmid specific proteins were labeled in minicells and analysed by SDS-PAGE. A 34,000 dalton molecular weight protein was identified to be missing in the amber mutant of plasmid mini-F. |
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