| First Author | Tam SY | Year | 1990 |
| Journal | Mol Cell Biol | Volume | 10 |
| Issue | 12 | Pages | 6619-23 |
| PubMed ID | 2174112 | Mgi Jnum | J:10836 |
| Mgi Id | MGI:59281 | Doi | 10.1128/mcb.10.12.6619 |
| Citation | Tam SY, et al. (1990) Functional expression of the genomic DNA sequences encoding mouse Na,K-ATPase alpha 1 gene by cotransfection of overlapping genomic DNA segments. Mol Cell Biol 10(12):6619-23 |
| abstractText | The 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. |
