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PMID:6345791

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Citation

Hanahan, D (1983) Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166:557-80

Abstract

Factors that affect the probability of genetic transformation of Escherichia coli by plasmids have been evaluated. A set of conditions is described under which about one in every 400 plasmid molecules produces a transformed cell. These conditions include cell growth in medium containing elevated levels of Mg2+, and incubation of the cells at 0 degrees C in a solution of Mn2+, Ca2+, Rb+ or K+, dimethyl sulfoxide, dithiothreitol, and hexamine cobalt (III). Transformation efficiency declines linearly with increasing plasmid size. Relaxed and supercoiled plasmids transform with similar probabilities. Non-transforming DNAs compete consistent with mass. No significant variation is observed between competing DNAs of different source, complexity, length or form. Competition with both transforming and non-transforming plasmids indicates that each cell is capable of taking up many DNA molecules, and that the establishment of a transformation event is neither helped nor hindered significantly by the presence of multiple plasmids.

Links

PubMed

Keywords

Cations; DNA, Superhelical; Dimethyl Sulfoxide/pharmacology; Dithiothreitol/pharmacology; Escherichia coli/drug effects; Escherichia coli/genetics; Escherichia coli/growth & development; Plasmids/drug effects; Species Specificity; Transformation, Bacterial/drug effects

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