Linked Life Data

2174112

Source:http://linkedlifedata.com/resource/pubmed/id/2174112

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pubmed-article:2174112pubmed:abstractTextThe entire 33-kb coding region of the mouse Na,K-ATPase alpha 1 subunit gene was cloned in two overlapping cosmids which contain inserts of 40 kb. To assess the functional expression of the mouse alpha 1 gene, the two cosmids were cotransfected into ouabain-sensitive CV-1 monkey cells yielding an average of 64 resistant colonies per 10(6) cells per microgram of DNA. Analysis of the DNA transferred to the ouabain-resistant transformants by the two cosmids suggests that the generation of a functional gene can occur by homologous recombination between the two introduced segments, as demonstrated by generation of a novel diagnostic restriction fragment. The ability to reconstruct the intact mouse alpha 1 gene in a heterologous host cell and to monitor its functional expression with a selection protocol permits direct identification and isolation of regulatory sequences for the gene.lld:pubmed
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pubmed-article:2174112pubmed:articleTitleFunctional expression of the genomic DNA sequences encoding mouse Na,K-ATPase alpha 1 gene by cotransfection of overlapping genomic DNA segments.lld:pubmed
pubmed-article:2174112pubmed:affiliationCenter for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.lld:pubmed
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pubmed-article:2174112pubmed:publicationTypeResearch Support, U.S. Gov't, P.H.S.lld:pubmed
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