Genetic analysis of two bacterial RNA polymerase mutants that inhibit the growth of bacteriophage T7

Sara R. Buchstein, David C. Hinkle

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10 Scopus citations

Abstract

The Escherichia coli mutants 7009 and BR3 are defective in the growth of bacteriophage T7. We have previously shown that both of these mutant hosts produce an altered RNA polymerase which is resistant to inhibition by the T7 gene 2 protein (De Wyngaert and Hinkle 1979). In both strains, the mutation which prevents T7 growth is closely linked to rifA (rpoB). Both mutants are complemented by transformation with a multicopy plasmid carrying rpoB and rpoC but not by a plasmid carrying only rpoB. This indicates that the mutations reside in rpoC, the structural gene for the β′ subunit of RNA polymerase. When a single copy of the wildtype rpoC allele is introduced into the mutant using the transducing phage λdri fd18, the mutant allele is dominant over wildtype. The λdri fd18 transductant also remains unable to support the growth of T7 in the presence of rifampin. This supports our conclusion that the mutation is in rpoC. We have measured the growth of T7 phage, the kinetics of phage DNA synthesis, and the structure of replicative DNA intermediates in several transductants, and compared these results with those obtained in the original mutant strains.

Original languageEnglish
Pages (from-to)211-218
Number of pages8
JournalMolecular Genetics and Genomics
Volume188
Issue number2
DOIs
StatePublished - Dec 1982
Externally publishedYes

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