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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1995-12-26
|
| pubmed:abstractText |
We report here the design, construction and testing of a self-inactivating (Sin) retrovirus promoter-trap vector suitable for identifying and isolating transcriptionally active regions from the mouse genome. When this vector, which contains the bacterial aph gene as its reporter, is integrated into a site downstream from an active host cell promoter, it expresses aph, whose product, aminoglycoside phosphotransferase, produces resistance to the antibiotic G418 in mammalian cells. The construct also contains a native aph promoter which functions in bacteria, but not in mouse cells, to express kanamycin (Km) resistance, plus an adjacent pBR322-derived replication origin. Thus, mammalian DNA segments containing actively transcribed regions flanking aph can be quickly isolated by restriction endonuclease treatment of total DNA from provirus-containing mouse cells, followed by self-ligation, transformation and Km selection of plasmids carried by bacteria transformed with this DNA. We tested this Sin retrovirus promoter-trap system by isolating eight DNA segments upstream to the provirus integration sites in the genome of virus-infected mouse F9 cells. We found that the Sin retrovirus vector produces a high yield of infectious virus particles carrying aph, and that the isolated genomic DNA fragments of F9 cells are transcriptionally active.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Neoplasm,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers,
http://linkedlifedata.com/resource/pubmed/chemical/Kanamycin Kinase,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphotransferases (Alcohol Group...
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Oct
|
| pubmed:issn |
0378-1119
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
27
|
| pubmed:volume |
164
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
289-94
|
| pubmed:dateRevised |
2008-11-21
|
| pubmed:meshHeading |
pubmed-meshheading:7590345-3T3 Cells,
pubmed-meshheading:7590345-Animals,
pubmed-meshheading:7590345-Base Sequence,
pubmed-meshheading:7590345-DNA,
pubmed-meshheading:7590345-DNA, Neoplasm,
pubmed-meshheading:7590345-DNA Primers,
pubmed-meshheading:7590345-Genetic Vectors,
pubmed-meshheading:7590345-Genome,
pubmed-meshheading:7590345-Kanamycin Kinase,
pubmed-meshheading:7590345-Mammals,
pubmed-meshheading:7590345-Mice,
pubmed-meshheading:7590345-Molecular Sequence Data,
pubmed-meshheading:7590345-Phosphotransferases (Alcohol Group Acceptor),
pubmed-meshheading:7590345-Plasmids,
pubmed-meshheading:7590345-Polymerase Chain Reaction,
pubmed-meshheading:7590345-Promoter Regions, Genetic,
pubmed-meshheading:7590345-Proviruses,
pubmed-meshheading:7590345-Restriction Mapping,
pubmed-meshheading:7590345-Retroviridae,
pubmed-meshheading:7590345-Teratocarcinoma,
pubmed-meshheading:7590345-Transcription, Genetic,
pubmed-meshheading:7590345-Tumor Cells, Cultured,
pubmed-meshheading:7590345-Virus Integration
|
| pubmed:year |
1995
|
| pubmed:articleTitle |
Rapid identification and isolation of transcriptionally active regions from mouse genomes.
|
| pubmed:affiliation |
Institute of Molecular Biology, Academia Sinica, Nankang Taipei, Taiwan, ROC.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|