人WDR79基因启动子表达载体的构建及鉴定

CONSTRUCTION AND IDENTIFICATION OF THE EXPRESSION VECTOR CONTAINING THE HUMAN WDR79 GENE PROMOTER

人WDR79是一种重要的支架蛋白,在端粒酶组装、卡哈尔体(Cahar body)形成和DNA损伤修复等过程中发挥着重要作用。采用PCR扩增技术获得人WDR79基因启动子上游DNA序列,构建人WDR79基因启动子荧光素酶报告基因载体pGL3-WDR79-promoter;通过双酶切、琼脂糖凝胶电泳、DNA测序和荧光素酶活性测定等实验手段,验证质粒pGL3-WDR79-promoter构建的正确性及其表达活性。本研究结果为进一步探讨人WDR79基因表达的调控机制奠定实验基础。

WDR79, a scaffold protein, plays an critical role in telomerase assembly, Cahar body formation, and DNA damage repair. The upstream DNA sequence of human WDR79 promoter was obtained by PCR amplification to construct the recombination vector pGL3-WDR79- promoter. The correctness and activity of the plasmid pGL3-WDR79-promoter were verified by double enzyme cutting, agarose gel electrophoresis, DNA sequencing and luciferase assasies. The results of this study provide experimental basis for elucidating the mechanism of WDR79 gene expression.