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Transformation in Bacillus subtilis

Fate of transforming DNA in transformation deficient mutants

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Summary

Transformation deficient mutants were isolated by means of selection for sensitivity to methylmethane-sulfonate (MMS). The mutations were introduced into a multiple auxotrophic highly transformable recipient. The transformation deficient strains were characterized with respect to their sensitivity to UV-irradiation and treatment with MMS and mitomycin-C (MC) and with respect to both the physico-chemical and biological properties of reextracted donor DNA.

As has been established previously (Davidoff-Abelson and Dubnau, 1973b) in the transformation proficient wild-type, double-stranded fragments (DSF), single-stranded fragments (SSF) and donorrecipient complex (DRC) are formed from donor DNA.

The mutants we report on are of various types: Mutant 7G-73 (transformation frequency about 25 times lower than in the wild-type) is sensitive to UV-irradiation and treatment with MMS and MC, and is extremely deficient in the production of DRC.

Mutant 7G-84 (transformation frequency about 12 times lower than in the wild-type) shows also sensitivity to UV-irradiation and treatment with MMS and MC. However, although it forms DSF, SSF, and DRC, the biological activity of DNA re-extracted from transforming cultures of 7G-84 is much reduced as compared to that of the wild-type.

Mutant 7G-97 (transformation frequency about 500 times lower than in the wild-type) shows approximately wild-type resistance to UV-irradiation and treatment with MMS and MC, and forms DSF exclusively; the donor DNA is not processed further.

Double mutants6G-103 and 6G-105, constructed by transformation of mutant 7G-97, with DNA from 7G-84 and 7G-73, are about 1250 and 5000 times less transformable than the wild-type respectively. They are sensitive to UV-irradiation and treatment with MMS and MC. Mutants 6G-103 and 6G-105 also produce DSF, which are not processed further.

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Authors and Affiliations

  1. Department of Genetics, Biological Centre, University of Groningen, Kerklaan 30, Haren (Gn), The Netherlands

    Johannes Buitenwerf & Gerard Venema

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  1. Johannes Buitenwerf
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  2. Gerard Venema
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Communicated by W. Arber

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Buitenwerf, J., Venema, G. Transformation in Bacillus subtilis . Molec. gen. Genet. 151, 203–213 (1977). https://doi.org/10.1007/BF00338696

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