论文部分内容阅读
目的 为消除急性髓系白血病标志基因AML1/ETO的活性 ,确定核受体辅助抑制因子(nuclearreceptorco repressor,N CoR)与ETO基因相互作用的活性部位。方法 用PCR扩增不同区段的N CoR(N CoRY) ,克隆入酵母表达质粒pGADGL中 ,构建获得pGADGL/N CoRY。应用酵母双杂交技术及X gal染色确定pGADGL/N CoRY 10个区段的N CoR是否与ETO结合。结果 N CoR的 988～ 112 6氨基酸残基与 15 5 1～ 180 1氨基酸残基共存时 ,才能与ETO发生结合作用 ,是与ETO相互作用的结构域。结论 N CoR两个结构域能与ETO的两个锌指结构作用 ,保持两种蛋白之间的稳定结合。
Objective To determine the activity of AML1/ETO, an acute myeloid leukemia marker gene, to determine the active site of interaction between nuclear receptor receptor repressor (N CoR) and ETO gene. Methods Different segments of N CoR (N CoRY) were amplified by PCR and cloned into yeast expression plasmid pGADGL to construct pGADGL/N CoRY. Yeast two-hybrid technology and X gal staining were used to determine whether NCoR of 10 segments of pGADGL/N CoRY was bound to ETO. Results The colocalization of 988 to 112 6 amino acid residues and 155 to 1801 amino acid residues of N CoR can interact with ETO and is a domain that interacts with ETO. Conclusion The two N CoR domains can interact with two zinc finger structures of ETO and maintain stable binding between the two proteins.