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Construction of spore mutants of Bacillus subtilis for the development as a host for foreign protein production

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Abstract

For the development of Bacillus subtilis as a host for foreign protein synthesis, three types of sigma factor deleted mutants (spoIIAC, spoIIIG and spoIIIC) were constructed by antibiotic marker insertion using plasmid vector-mediated method or LFH (Long Flanking Homology)-PCR. Mother cell specific sigma factor mutants of B. subtilis (σK), B. subtilis DB104 ΔspoIIIC (km r)::pMK101, had two to three times higher subtilisin activity than the wild type DB104::pMK101. Subtilisin expression by the other two mutants, B. subtilis DB104 ΔspoIIAC (km r)::pMK101 and DB104 ΔspoIIIG (km r)::pMK101, which are pre-spore specific sigma factor (σF and σG) deleted strains, was similar to, or less than that of the wild type.

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Authors and Affiliations

  1. Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University, Seoul, 151-742, Korea

    June-Hyung Kim & Byung-Gee Kim

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  1. June-Hyung Kim
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  2. Byung-Gee Kim
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Correspondence to Byung-Gee Kim.

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Kim, JH., Kim, BG. Construction of spore mutants of Bacillus subtilis for the development as a host for foreign protein production. Biotechnology Letters 23, 999–1004 (2001). https://doi.org/10.1023/A:1010515632509

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  • DOI: https://doi.org/10.1023/A:1010515632509

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