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Temperature sensitive mutations affecting RNA synthesis in Escherichia coli

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Summary

A streptomycin method has been used for the isolation of mutants with RNA synthesis inhibited at elevated temperature. The method is based on the observation that streptomycin kills bacteria with normal RNA synthesis and does not affect the cells with inhibited synthesis of RNA. This selection method increases the yield of temperature sensitive mutants by a factor 10–20, the amount of mutants with disturbed RNA synthesis is increased 3–5 fold as compared with the method of replicas.

Several types of mutants were found among the temperature sensitive strains: those possessing temperature sensitivity of one, two or three types of cellular macromolecules DNA, RNA and protein. The screening among the mutants with affected RNA synthesis revealed a strain ts-19 showing low RNA polymerase activity in cell extracts and partially purified RNA polymerase preparations. The presented evidence suggests that ts-19 mutation affects the structural gene of one of the RNA polymerase subunits.

The mapping of the corresponding locus indicated that it was located between the str and thy loci in E. coli K 12 chromosome at a distance of about 20 recombination units from the first locus.

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Authors and Affiliations

  1. Kurchatov Institute of Atomic Energy, Moscow

    R. B. Khesin, S. Z. Mindlin, Zh. M. Gorlenko & T. S. Ilyina

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  1. R. B. Khesin
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  2. S. Z. Mindlin
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  3. Zh. M. Gorlenko
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  4. T. S. Ilyina
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Communicated by D. Goldfarb

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Khesin, R.B., Mindlin, S.Z., Gorlenko, Z.M. et al. Temperature sensitive mutations affecting RNA synthesis in Escherichia coli . Molec. Gen. Genet. 103, 194–208 (1968). https://doi.org/10.1007/BF00427146

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  • DOI: https://doi.org/10.1007/BF00427146

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