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Transformation of Bacillus licheniformis protoplasts by plasmid DNA

Department of biotechnology, Gödöllö University of Agricultural Sciences, Gödöllö, Páter u. 1., H-2103, Hungary

ABSTRACT

Summary: A simple and efficient protoplast regeneration and PEG-induced transformation procedure was developed for Bacillus licheniformis. The protoplast stabilizing osmoticum was 0.5 M sucrose in the regenerating media, and the restoration of cell wall was carried out at pH 8.4. The mean frequency of regeneration was 83%. The transformation efficiency was studied with four different strains and five plasmids, and ranged from 3.5x10⁵to 7.2x10⁶transformants per g plasmid DNA depending on the plasmid, and on the donor and recipient strains used. Endonuclease digested and ligated plasmid DNA could also be used to transform the protoplasts, but at a lower frequency.

Author for correspondence: Zoltán Prágai. Fax: +36 28 30804. e-mail: pz@rlt.gau.hu.