蛋白磷酸酶2A Cα在昆虫杆状病毒中的表达及活性

摘要

目的 利用昆虫杆状病毒系统表达蛋白磷酸酶(PP)2A Cα亚基并进行活性分析.方法 先将PP2A Cα构建进入昆虫杆状病毒表达载体,然后将其转染到昆虫细胞Sf9中并进行蛋白表达,最后检测PP2A Cα的功能.结果 Western印迹结果表明PP2A Cα蛋白已经在昆虫细胞成功表达;PP2A活性检测和GST pull-down实验结果表明PP2A Cα蛋白既有生物学功能,又有结构学功能.结论 昆虫杆状病毒载体系统可以成功表达有活性的PP2A Cα蛋白.%Objective To construct the recombinant baculovirusof protein phosphatase 2A(PP2A)Cα and express the PP2A Cα protein in insect cells.Methods Firstly,the gene of PP2A Cα was cloned into pFastBac and transpositioned into the genome of insect baculovirus.Then, the PP2A Cα protein was expressed in the insect Sf9 cells by the recombinant baculovirus.Finally, PP2A Cα activity was assayed.Results The protein of PP2A Cα was successfully expressed in the insect cells and it showed its biological activity.Conclusions The insect baculovirus could be used to express the PP2A Cα protein.