Nonreciprocal exchanges of information between DNA duplexes coinjected into mammalian. cell nuclei

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Publication Type Journal Article
School or College College of Science; School of Medicine
Department Internal Medicine; Oncological Sciences; Human Genetics; Biology
Program Institute of Human Genetics; Howard Hughes Medical Institute (HHMI)
Creator Thomas, Kirk R.; Capecchi, Mario R.
Other Author Folger, Kim R.
Title Nonreciprocal exchanges of information between DNA duplexes coinjected into mammalian. cell nuclei
Date 1985-01
Description We have examined the mechanism of homologous recombination between plasmid molecules coinjected into cultured mammalian cells. Cell lines containing recombinant DNA molecules were obtained by selecting for the reconstruction of a functional Neor gene from two plasmids that bear different amber mutations in the Neor gene. In addition, these plasmids contain restriction-length polymorphisms within and near the Neor gene. These polymorphisms did not confer a selectable phenotype but were used to identify and categorize selected and nonselected recombinant DNA molecules. The striking conclusion from this analysis is that the predominant mechanism for the exchange of information between coinjected plasmid molecules over short distances (i.e., less than 1 kilobase) proceeds via nonreciprocal homologous recombination. The frequency of homologous recombination between coinjected plasmid molecules in cultured mammalian cells is extremely high, approaching unity. We demonstrate that this high frequency requires neither a high input of plasmid molecules per cell nor a localized high concentration of plasmid DNA within the nucleus. Thus, it appears that plasmid molecules, once introduced into the nucleus, have no difficulty seeking each other out and participating in homologous recombination even in the presence of a vast excess of host DNA sequences. Finally, we show that most of the homologous recombination events occur within a 1-h interval after the introduction of plasmid DNA into the cell nucleus.
Type Text
Publisher American Society for Microbiology
Volume 5
Issue 1
First Page 59
Last Page 69
Subject Animals; Cells, Cultured; DNA Restriction Enzymes; Kinetics
Subject MESH Recombination, Genetic; Plasmids; DNA
Language eng
Bibliographic Citation Mol Cell Biol. 1985 Jan;5(1):59-69: Folger KR, Thomas KR. Capecchi MR. Nonreciprocal exchanges of information between DNA duplexes coinjected into mammalian. cell nuclei. Retrieved on September 15.2006 from http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=2984556
Rights Management Copyright 1985 by American Society for Microbiology
Format Medium application/pdf
Identifier ir-main,413
ARK ark:/87278/s6611ht2
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2012-06-13
ID 705996
Reference URL https://collections.lib.utah.edu/ark:/87278/s6611ht2
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