1887

Abstract

Methods have been developed for chemical transformation and electro-transformation (electroporation) of vegetative cells of 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 transformant c.f.u. per viable c.f.u. (1 × 10 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 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 c.f.u. per viable c.f.u. (2.6 × 10 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.

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1990-07-01
2021-10-16
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