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Studies of transformation of Escherichia coli with plasmids (1983)
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| Venue: | J. Mol. Biol |
| Citations: | 1609 - 1 self |
BibTeX
@ARTICLE{Hanahan83studiesof,
author = {Douglas Hanahan},
title = {Studies of transformation of Escherichia coli with plasmids},
journal = {J. Mol. Biol},
year = {1983},
pages = {557--580}
}
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Abstract
Factors that affect he probability of genetic transformation f 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 Mg 2+. and incubation of the cells at 0 ~ in a solution of Mn 2+, ("a 2+, Rb + or K +, dimethyl sulfoxide, dithiothreitol, and hexamine cobalt (III). Transibrmation efficiency declines linearly with increasing plasmid size. Relaxed and supercoiled plasmids transfol'm with similar probabilities. Non-transforming DNAs compete consistent with mass. No significant variation is observed between competing DNAs of difi~rent 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. 1. Introduct ion Both gram-posit ive and gram-negative bacteria can take up and stably establish
Keyphrases
escherichia coli cell growth plasmid transfol multiple plasmid transformation event gram-negative bacteria hexamine cobalt difi rent source non-transforming dna compete consistent plasmid molecule many dna molecule dimethyl sulfoxide gram-posit ive non-transforming plasmid genetic transformation escherichia coli introduct ion transformed cell significant variation medium containing elevated level plasmid size transibrmation efficiency similar probability
