RRC ID 30081
Author Hanahan D.
Title Studies on transformation of Escherichia coli with plasmids.
Journal J Mol Biol
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.
Volume 166(4)
Pages 557-80
Published 1983-6-5
DOI 10.1016/s0022-2836(83)80284-8
PII S0022-2836(83)80284-8
PMID 6345791
MeSH 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
IF 5.067
Times Cited 9216
Prokaryotes E. coli ME9033