Linked Life Data

16674971

Source:http://linkedlifedata.com/resource/pubmed/id/16674971

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-2
pubmed:dateCreated
2006-6-5
pubmed:abstractText
Tamoxifen treatment allows MerCreMer fusion recombinase to localize to the nucleus where MerCreMer can excise a floxed inhibitory DNA segment, thereby activating the expression of a downstream gene. This excision is irreversible, and it is therefore difficult to predict which non-activated clones will express the gene at high levels after recombination. We transfected a vector using HLA-A2.1 as floxed inhibitory DNA element and its expression level as surrogate marker predicting future expression of the attenuated downstream target gene. The target gene encoded an EGFP-linked fusion protein. In the unsorted population, 6% of the cells expressed the transfected target gene after recombination and less than 10-fold higher than the population before recombination. However after flow-cytometric selection for high HLA-A2.1 expression, 47% of the cells expressed the target gene after recombination and at levels 37-fold higher than the sorted population before recombination. 58% of the clones were capable of expressing the fusion protein and some over 200-fold above background of untransfected cells and greater than 20-fold higher levels of expression than before recombination. We describe an efficient method to select for clones expressing high levels of a target gene after tamoxifen regulated Cre-loxP recombination. Other floxed reporter genes should be equally useful.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0022-1759
pubmed:author
pubmed:issnType
Print
pubmed:day
30
pubmed:volume
312
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
201-8
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
2006
pubmed:articleTitle
Floxed reporter genes: Flow-cytometric selection of clonable cells expressing high levels of a target gene after tamoxifen-regulated Cre-loxP recombination.
pubmed:affiliation
Department of Pathology, The University of Chicago, 5831 South Ellis Avenue, Chicago, IL 60637, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural