Linked Life Data

8254657

Source:http://linkedlifedata.com/resource/pubmed/id/8254657

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1994-1-12
pubmed:abstractText
The FLP site-specific recombinase is encoded by the two micron circle, an endogenous plasmid of Saccharomyces cerevisiae. FLP-mediated recombination in vitro proceeds via a short-lived Holliday (chi) intermediate. We have made a synthetic chi structure containing two FLP recognition target (FRT) sequences in order to investigate resolution by purified FLP protein. We found that incubation of this model substrate with FLP generated two pairs of linear products in equal quantities. Thus, resolution was equally likely to occur in either direction. Alteration of FLP binding sites, so as to inhibit binding, affected the direction of resolution; cleavage was reduced adjacent to the altered binding site. The overall efficiency of resolution increased when one FLP binding site was mutated. In investigating the series of mutated chi structures we found that resolution requires only two intact FLP binding sites. However, the non-specific protein-DNA interaction of additional FLP molecules may also be required. Thus, resolution is more tolerant of the loss of FLP binding sites than is the complete recombination reaction.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0022-2836
pubmed:author
pubmed:issnType
Print
pubmed:day
5
pubmed:volume
234
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
522-33
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1993
pubmed:articleTitle
Resolution of synthetic chi structures by the FLP site-specific recombinase.
pubmed:affiliation
Department of Molecular and Medical Genetics, University of Toronto, Ontario, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't