克隆鸡胸苷激酶基因的突变分析。

T J Kwoh, D Zipser, M Wigler
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引用次数: 0

摘要

我们分析了鸡胸苷激酶(tk)基因的转录和编码单元。采用Heffron等人的体外连接子插入法构建了鸡tk突变基因文库。在包含该基因的3.0 kbp HindIII片段中,共分离并定位了125个突变。每个突变对胸苷激酶基因的影响是通过小鼠Ltk细胞转染效率的测量来确定的。鸡的tk mRNA约为2 kb,并被聚腺苷化。转录方向也被确定。根据这些结果,我们提出了一种基因结构,其中至少有三个小内含子将氨基酸编码区分隔成至少四个片段。
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Mutational analysis of the cloned chicken thymidine kinase gene.

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.

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