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目的:检测KSHV编码的vFLIP蛋白对Jurkat细胞HIV抑制因子表达的影响。方法:本研究首先构建真核表达载体pIRES2-EGFP-vFLIP,再将其电穿孔至Jurkat细胞,然后使用实时荧光定量PCR技术检测Jurkat细胞中A3G、A3F、CCL5等HIV抑制因子的表达水平,从而探讨vFLIP抗HIV/AIDS的作用及机制。结果:成功构建了真核表达载体pIRES2-EGFP-vFLIP,电转染效率达到40%左右。与对照载体转染组相比,vFLIP转染组内Jurkat细胞中CCL5、A3G、A3F及Mx2基因的表达分别上调了6.71、2.20、1.52及14.47倍。结论:vFLIP蛋白可以激活Jurkat细胞内某些HIV抑制因子的表达,提示其具有一定的抗HIV/AIDS作用。
Objective: To investigate the effect of vFLIP protein encoded by KSHV on the expression of HIV inhibitor in Jurkat cells. Methods: The eukaryotic expression vector pIRES2-EGFP-vFLIP was constructed and then electroporated into Jurkat cells. Real-time fluorescent quantitative PCR was used to detect the expression of HIV inhibitors such as A3G, A3F and CCL5 in Jurkat cells To investigate the anti-HIV / AIDS effect and mechanism of vFLIP. Results: The eukaryotic expression vector pIRES2-EGFP-vFLIP was successfully constructed and the efficiency of electroporation was about 40%. Compared with the control vector transfection group, the expression of CCL5, A3G, A3F and Mx2 genes in Jurkat cells in vFLIP transfected group were up-regulated 6.71, 2.20, 1.52 and 14.47 fold, respectively. CONCLUSION: vFLIP protein can activate certain HIV inhibitors in Jurkat cells, suggesting that it has some anti-HIV / AIDS effects.