辐射敏感细胞ku80基因突变及其DNA结合活性研究

ku80 Gene Mutation of Radiosensitive Cell Line and Its DNA Binding Activity

RT PCR克隆辐射敏感细胞及其亲本细胞的ku80基因cDNA ,发现辐射敏感细胞的ku80基因与双链断裂DNA末端相互作用的位置存在基因突变 ,用凝胶阻滞和DNA 蛋白质印迹进一步证实突变ku基因编码蛋白结合双链断裂末端DNA的能力下降 。

ku80 gene of radiosensitive cell line SX 9 was cloned by RT PCR.Compared cDNA of the radiosensitive cell line with its parental cell line, it was found that there were two mutation sites in ku80 gene which could interact with DSB DNA and Ku70.It was confirmed by EMSA and Southwestern blot that the DSB end binding activity of Ku80 in the nucleus extract of the radiosensitive cell decreased greatly,which infered that the radiosensitivity of the cell line is caused by dysfunction of its Ku80 protein.