果蝇S2细胞中高表达荧光素酶重组质粒的构建及活性测定  

Construction of recombinant plasmid highly expressing luciferase in Drosophila S2 cells

作　　者：丁洁[1] 周静[1] 袁榴娣[1] 

机构地区：[1]东南大学基础医学院遗传与发育生物学系,江苏南京210009

出　　处：《东南大学学报（医学版）》2006年第6期403-406,共4页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(30270309)

摘　　要：目的:构建果蝇S2细胞中高表达荧光素酶的重组质粒。方法:用PCR方法扩增得到pac启动子的基因片段,经酶切亚克隆至pGL3-Basic的真核表达载体中,转染果蝇S2细胞,通过测定荧光素酶和β-半乳糖苷酶的活性计算荧光素酶的相对活性。结果:构建的重组质粒在S2细胞中荧光素酶的相对活性较pGL3-Control中增加了2.7万倍。结论:成功构建了果蝇S2细胞中高表达荧光素酶的重组质粒pGL3-pac。Objective To construct recombinant plasmid highly expressing luciferase in Drosophila S2 cells. Methotis Recombinant plasmid pGLa-pac was constructed by PCR and subcloned and transiently transfected into Drosophila S2 cells. The activity of LUC and fl-gal were determined to calculate the RLUC after transfection. Result Compared with pGLa-Control ,there was nearly 27 thousand increase in pGLa-pac activity. Conclusion The recombinant plasmid highly expressing luciferase has been successfully constructed in Drosophila S2 cells.

关 键 词：果蝇S2细胞 荧光素酶 启动子 报告基因 转染 

分 类 号：Q782[生物学—分子生物学]