Functional expression of the genomic DNA sequences encoding mouse Na,K-ATPase alpha 1 gene by cotransfection of overlapping genomic DNA segments.

S Y Tam; E N Geissler; S L Graw; D E Housman

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Functional expression of the genomic DNA sequences encoding mouse Na,K-ATPase alpha 1 gene by cotransfection of overlapping genomic DNA segments.

机译：通过共转染重叠的基因组DNA片段，编码小鼠Na，K-ATPase alpha 1基因的基因组DNA序列的功能表达。

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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.

机译：将小鼠Na，K-ATP酶α1亚基基因的整个33kb编码区克隆到两个重叠的粘粒中，所述粘粒包含40kb的插入片段。为了评估小鼠alpha 1基因的功能表达，将这两种粘粒共转染到对哇巴因敏感的CV-1猴细胞中，每10微克DNA中平均每10（6）个细胞产生64个抗性菌落。对通过两个粘粒转移至哇巴因抗性转化子的DNA的分析表明，可以通过两个引入的片段之间的同源重组来发生功能基因的产生，如通过产生新的诊断性限制性片段所证实的。在异源宿主细胞中重建完整小鼠alpha 1基因并通过选择方案监控其功能表达的能力可直接鉴定和分离该基因的调控序列。

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《Molecular and Cellular Biology》 |1990年第12期|共5页
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S Y Tam; E N Geissler; S L Graw; D E Housman;
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中图分类细胞生物学;
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6. Functional expression of the genomic DNA sequences encoding mouse NaK-ATPase alpha 1 gene by cotransfection of overlapping genomic DNA segments. [O] . S Y Tam, E N Geissler, S L Graw, 1990

机译：通过共转染重叠的基因组DNA片段编码小鼠NaK-ATPase alpha 1基因的基因组DNA序列的功能表达。
7. Functional expression of the genomic DNA sequences encoding mouse Na,K-ATPase alpha 1 gene by cotransfection of overlapping genomic DNA segments. [O] . Tam, S Y, Geissler, E N, Graw, S L, 1990

机译：通过共转染重叠的基因组DNA片段，编码小鼠Na，K-ATPase alpha 1基因的基因组DNA序列的功能表达。

Functional expression of the genomic DNA sequences encoding mouse Na,K-ATPase alpha 1 gene by cotransfection of overlapping genomic DNA segments.

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