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目的 克隆ATM全长cDNA及含特异功能域的cDNA片段 ,寻找与ATM相互作用的蛋白 ,分析ATM在DNA损伤监视中的分子机理。方法 利用长片段PCR扩增法 ,从人外周血来源的cDNA库中扩增ATMcDNA ;利用酵母双杂交系统筛选人外周血来源的cDNA库中与ATM相互作用的蛋白。结果 经重叠PCR扩增到ATM全长cDNA ,并筛选到数条与ATMPI3K激酶区相互作用的cDNA ,并对其序列进行分析。结论 ATM通过与多种蛋白相互作用 ,传递DNA损伤信号
Objective To clone full-length cDNA of ATM and cDNA fragments containing specific domains to find ATM-interacting proteins and analyze the molecular mechanism of ATM in DNA damage surveillance. Methods The cDNA fragments of ATMcDNA were amplified from the cDNA library of human peripheral blood by using the method of long fragment PCR. The yeast two - hybrid system was used to screen the proteins interacting with ATM in cDNA library of human peripheral blood. Results The full-length cDNA of ATM was amplified by overlap PCR and several cDNAs interacting with ATMPI3K kinase were screened and their sequences were analyzed. Conclusion ATM interacts with many proteins to transmit DNA damage signals