Linked Life Data

8804858

Source:http://linkedlifedata.com/resource/pubmed/id/8804858

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1996-12-9
pubmed:abstractText
One of the challenges in human genome research is to identify the promoter sequences which play a key role in the regulation of gene expression. We report here a new promoter trapping system for use with mammalian cells comprised of the following three steps: 1) Cloning of DNA fragments into a promotertrapping vector, 2) integration of the trapping vector into a designated target in the mammalian genome using the Cre site-specific recombinase, and 3) screening of integrants for trapped promoter sequences by activation of the luciferase gene. To assess the efficiency of this system, lox trapping vectors containing sense tk promoter, antisense tk promoter, or a non-promoter sequence of the neo gene were employed. The resulting levels of luciferase activity of the site-specific integrants were measured directly. Luciferase activity of the integrants can be assayed under conventional culture conditions by simply replacing the culture medium with potassium phosphate buffer containing luciferin. Only those G418r colonies carrying the tk promoter in the normal orientation exhibited a 21-to 35-fold increase in luciferase activity over that of the other integrants. These results indicate that this system is an effective means of trapping promoter sequences from random mammalian genomic DNA fragments.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
1340-2838
pubmed:author
pubmed:issnType
Print
pubmed:day
30
pubmed:volume
3
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
73-80
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed:year
1996
pubmed:articleTitle
Trapping of mammalian promoters by Cre-lox site-specific recombination.
pubmed:affiliation
Ikeda GenoSPHERE Project, Tokai University School of Medicine, Kanagawa, Japan.
pubmed:publicationType
Journal Article