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-东南大学学报医学版

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

单位：东南大学基础医学院,遗传与发育生物学系,江苏,南京,210009

分类号：Q782

出版年·卷·期（页码）：2006·25·第六期（403-406）

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.Methods Recombinant plasmid pGL_3-pac was constructed by PCR and subcloned and transiently transfected into Drosophila S2 cells.The activity of LUC and β-gal were determined to calculate the RLUC after transfection.Result Compared with pGL_3-Control,there was nearly 27 thousand increase in pGL_3-pac activity.Conclusion The recombinant plasmid highly expressing luciferase has been successfully constructed in Drosophila S2 cells.

参考文献：

[1] LOUISE H. Reporter gene technology:the future looks bright. 1999. doi:10.1016/S0006-2952(99)00096-9
[2] GAMBHIR S S, BARRIO J R, HERSCHMAN H R. Assays for noninvasive imaging of reporter gene expression. 1999. doi:10.1016/S0969-8051(99)00021-9
[3] LECLERC G M, BOOCKFOR F R, FAUGHT W J. Development of a destabilized firefly luciferase enzyme for measurement of gene expression, 2000
[4] ARNOSTI D N. Design and function of transcriptional switches in Drosophila. 2002
[5] LERMAN D N, FEDER M E. Naturally occurring transposable elements disrupt hsp70 promoter function in Drosophila melanogaster. 2005(3). doi:10.1093/molbev/msi063

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