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1988•FEMS Microbiology LettersOpen access

Improvements in the transformation of Bacillus subtilis protoplasts with plasmid DNA

Clara P. Rubinstein, Laura C. Moratinos, Omar A. Coso, Carmen Sánchez-Rivas

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Abstract

The transformation system currently used for Bacillus subtilis protoplasts has been improved. Special emphasis was made on three parameters of practical importance: (a) conditions for direct selection of transformants, (b) optimization of the transformation system for Rec− mutants, and (c) conservation of protoplast suspensions for further use. Selective regeneration was efficiently achieved for kanamycin or neomycin. Chloramphenicol, tetracycline and erythromycin were only expressed when low concentrations of the antibiotics were used to select transformants during regeneration.

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The transformation system currently used for Bacillus subtilis protoplasts has been improved. Special emphasis was made on three parameters of practical importance: (a) conditions for direct selection of transformants, (b) optimization of the transformation system for Rec− mutants, and (c) conservation of protoplast suspensions for further use. Selective regeneration was efficiently achieved for kanamycin or neomycin. Chloramphenicol, tetracycline and erythromycin were only expressed when low concentrations of the antibiotics were used to select transformants during regeneration.

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Available abstract

The transformation system currently used for Bacillus subtilis protoplasts has been improved. Special emphasis was made on three parameters of practical importance: (a) conditions for direct selection of transformants, (b) optimization of the transformation system for Rec− mutants, and (c) conservation of protoplast suspensions for further use. Selective regeneration was efficiently achieved for kanamycin or neomycin. Chloramphenicol, tetracycline and erythromycin were only expressed when low concentrations of the antibiotics were used to select transformants during regeneration.

Key concepts: Bacillus subtilis, Plasmid, Protoplast, Transformation (genetics), Bacillaceae, DNA, Microbiology, Bacillales

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