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利用PCR方法对锌指蛋白ZNF191(243-368)的Ⅱ323和Arg327进行定点突变,成功地获得了突变体蛋白ZNF191(243～368)1323W和R327W;并利用荧光光谱研究了锌指蛋白和它的突变体蛋白ZNF191(243-368)1323W和ZNF191(243-368)R327W与寡聚核苷酸的相互作用.以溴化乙锭(EB)为荧光探针,考察了1323W和R327W的突变对锌指蛋白ZNF191(243～368)结合寡聚核苷酸的影响.并计算了锌指蛋白突变体1323W和R327W与DNA的结合常数Kz.讨论了可能的结合模式.
The mutants ZNF191 (243 ~ 368) 1323W and R327W were successfully obtained by site-directed mutagenesis of Ⅱ323 and Arg327 from zinc finger protein ZNF191 (243-368) by PCR. Fluorescence spectra of zinc finger protein and its Interaction of mutant proteins ZNF191 (243-368) 1323W and ZNF191 (243-368) R327W with oligonucleotides.Using ethidium bromide (EB) as a fluorescent probe, the effects of mutation of 1323W and R327W on the activity of zinc Refers to the effect of binding protein ZNF191 (243-368) to oligonucleotides, and the binding constants Kz of zinc finger protein mutants 1323W and R327W to DNA were calculated. Possible binding modes were discussed.