
Full text loading...
Methods have been developed for chemical transformation and electro-transformation (electroporation) of vegetative cells of Bacillus anthracis with supercoiled plasmid DNA. Chemical transformation was dependent on incubation in Tris/HCl with osmotic support and transformation with plasmid DNA was effected by treatment with polyethylene glycol 3350. Maximum transformation frequencies were 3·8 × 10−5 transformant c.f.u. per viable c.f.u. (1 × 103 c.f.u. per g DNA). Optimal frequencies were pH dependent and were affected by growth-medium composition. Transformation was not observed with linear or multimeric plasmid DNA. Electro-transformation of B. anthracis using high field intensity electroporation was dependent on the composition of both the growth medium and the electroporation buffer. Maximum electro-transformation frequencies were 5·3 × 10−4 c.f.u. per viable c.f.u. (2.6 × 104 c.f.u. per g DNA). The use of early exponential phase cells was critical to both procedures and the maximum efficiency (c.f.u. per g DNA) of each system was strain dependent under the conditions described.
Article metrics loading...
Full text loading...
References
Data & Media loading...