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克隆的鸡胸苷激酶基因的突变分析。

Mutational analysis of the cloned chicken thymidine kinase gene.

作者信息

Kwoh T J, Zipser D, Wigler M

出版信息

J Mol Appl Genet. 1983;2(2):191-200.

PMID:6875426
Abstract

We have analyzed the transcription and coding unit of the chicken thymidine kinase (tk) gene. We have constructed a library of mutant chicken tk genes by the in vitro linker insertion method of Heffron et al. A total of 125 mutations within a 3.0 kbp HindIII fragment containing the gene were isolated and mapped. The effect of each mutation upon the thymidine kinase gene was determined by measurement of the transfection efficiency in mouse Ltk- cells. The chicken tk mRNA is about 2 kb and polyadenylated. The direction of transcription was also determined. From these results, we propose a structure for the gene in which at least three small introns separate the amino acid coding region into at least four segments.

摘要

我们分析了鸡胸苷激酶(tk)基因的转录和编码单元。我们通过Heffron等人的体外接头插入法构建了一个突变鸡tk基因文库。在包含该基因的3.0kbp HindIII片段内共分离并定位了125个突变。通过测量在小鼠Ltk-细胞中的转染效率来确定每个突变对胸苷激酶基因的影响。鸡tk mRNA约为2kb且带有多聚腺苷酸。还确定了转录方向。根据这些结果,我们提出了该基因的一种结构,其中至少三个小内含子将氨基酸编码区域分隔成至少四个区段。

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