Access

You are not currently logged in.

Access your personal account or get JSTOR access through your library or other institution:

login

Log in to your personal account or through your institution.

If you need an accessible version of this item please contact JSTOR User Support

Site-Specific DNA Recombination in Mammalian Cells by the Cre Recombinase of Bacteriophage P1

Brian Sauer and Nancy Henderson
Proceedings of the National Academy of Sciences of the United States of America
Vol. 85, No. 14 (Jul. 15, 1988), pp. 5166-5170
Stable URL: http://www.jstor.org/stable/32380
Page Count: 5
  • Read Online (Free)
  • Cite this Item
If you need an accessible version of this item please contact JSTOR User Support
Site-Specific DNA Recombination in Mammalian Cells by the Cre Recombinase of Bacteriophage P1
Preview not available

Abstract

The Cre protein encoded by the coliphage P1 is a 38-kDa protein that efficiently promotes both intra- and intermolecular synapsis and recombination of DNA both in Escherichia coli and in vitro. Recombination occurs at a specific site, called lox, and does not require any other protein factors. The Cre protein is shown here also to be able to cause synapsis of DNA and site-specific recombination in a mammalian cell line. A stable mouse cell line was established that expresses the Cre protein under the control of the Cd2+-inducible metallothionein I gene promoter. DNA recombination was monitored with DNA substrates containing two directly repeated lox sites. One such substrate is a circular plasmid with two directly repeated lox sites (lox2) flanking a marker gene and was introduced into cells by Ca3(PO4)2 transformation. As a second substrate we used a pseudorabies virus (a herpesvirus) containing a lox2 insertion designed to provide a sensitive detection system for recombination. In both cases, site-specific recombination in vivo is dependent on the presence of the Cre protein and occurs specifically at the 34-base-pair lox sites. These results demonstrate the controlled site-specific synapsis of DNA and recombination by a prokaryotic protein in mammalian cells and suggest that Cre-mediated site-specific recombination may be a useful tool for understanding and modulating genome rearrangements in eukaryotes.

Page Thumbnails

  • Thumbnail: Page 
5166
    5166
  • Thumbnail: Page 
5167
    5167
  • Thumbnail: Page 
5168
    5168
  • Thumbnail: Page 
5169
    5169
  • Thumbnail: Page 
5170
    5170