Transformation of Bacillus thuringiensis protoplasts by plasmid deoxyribonucleic acid

A method has been developed to transform plasmid deoxyribonucleic acid into protoplasts of the insect pathogen Bacillus thuringiensis. Protoplasts were formed by treatment of cells with lysozyme. The efficiency of formation of protoplasts was affected by the strain, the media, and the cell density. Deoxyribonucleic acid uptake was induced by polyethylene glycol. Deoxyribonucleic acid from the Staphylococcus aureus plasmid pC194 was used for transformation. Although this plasmid could not be isolated as a stable extrachromosomal element, its chloramphenicol resistance was transferred to the recipient protoplasts. This was confirmed by assay for the enzyme chloramphenicol acetyltransferase, which confers resistance to chloramphenicol. This suggested that pC194 acts as an insertion element in B. thuringiensis.