pEGFP-ORF2质粒在Vero细胞中的有效表达

pEGFP-ORF2 effective expression in Vero cells

目的:确立pEGFP-ORF2质粒在不同细胞密度,不同转染时间,及血清存在与否时在细胞中的有效表达,为进一步研究HSV-2LATORF2对细胞的作用,及阐明HSV-2潜伏复发机制打下基础。方法:采用脂质体法将pEGFP-ORF2与pEGFP-C2两个真核表达载体转染Vero细胞,观察荧光蛋白表达。结果:有无血清对转染效果无明显差异。在细胞密度为4.0×104cells/ml,转染后84hEGFP表达量较其它条件下高,且在相同条件下pEGFP-ORF2与pEGFP-C2的表达无明显差异。结论:血清对表达效果的影响不大,细胞的接种密度及转染时间对转染有明显作用,以荧光蛋白作为标签鉴定蛋白表达的方法切实可行。

Objective： To establish the condition ofpEGFP-ORF2 for effective expression in vero cells in different cell densities, transfected time and with or without serum, which can be used for explore HSV-2 LAT ORF2 regulation role on Vero cells and establish basis for the mechanisim studies of HSV-2 latency and reactivation.Methods The recombination vector pEGFP-ORF2 and pEGFP-C2 was transfected into Vero cells by liposome transfection reagent, EGFP-ORF2 was observed with green fluorescent protein.Results There was no difference in transfection with or without serum. When the cell density is 4.0×10^4 cells/ml and post-transfected 84 h later, the EGFP is an effective expressed. And in the same conditions, there is no different expression level between pEGFP-ORF2 and pEGFP-C2. Conclusion Serum has little effect on expression level. The cells density and transfection time play an importmant role .The methods is feasible to identify the expression ofHSV-2 LAT protein by green fluorescent protein tag.