染色质免疫沉淀技术及其在基因表达调控研究中的初步应用

Chromatin Immunoprecipitation and Its Preliminary Application for Gene Regulation

目的验证抑癌基因表达产物p53蛋白在体内是否结合于p21WAF1/CIP1基因的启动子区,并检测p53蛋白对热休克蛋白hsp90β基因启动子区的结合情况。方法采用染色质免疫沉淀(ChIP)及PCR技术检测特异基因调节序列,Western印迹分析p53蛋白。结果在p53抗体免疫沉淀的DNA片段中含有p21WAF1/CIP1和hsp90β基因的启动子区p53蛋白结合序列,免疫沉淀样品中含有p53蛋白。结论p53蛋白在体内结合于p21WAF1/CIP1和hsp90β基因的启动子区,参与两基因的表达调控。

Objective To verify the binding of p53to p21 WAF1/CIP1 gene promoter and detect its binding to hsp90βgene promoter in vivo.Methods Chromatin immunoprecipitation and PCR analysis were used to measure specific gene regulation sequence and Western blot analysis to investigate p53protein.Results The p53binding sequences on the promoters of p21 WAF1/CIP1 and hsp90βgene were found in the p53antibody immunoprecipitated DNA fragments and p53was detected in the immunoprecipitated samples.Conclusions p53binds to promoters of p21 WAF1/CIP1 and hsp90βgene in viv o,and regulate s the expression of the two genes.