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将丙肝病毒C +E1区基因插入绿色荧光报告基因 pEGFP N1中 ,构建真核表达重组质粒 pEGFP N1 HCV/C +E1。转染小鼠骨髓瘤细胞SP2 / 0 ,在荧光显微镜下观察绿色荧光融合蛋白的表达情况 ,结果在细胞浆中出现了绿色荧光 ,表明目的基因得到表达。再通过G418筛选后大量培养用作细胞毒实验的靶细胞 ,结果表明以EGFP报告基因作筛选标记制备的靶细胞完全可以满足细胞毒实验要求
The C + E1 region of hepatitis C virus was inserted into green fluorescent reporter gene pEGFP N1 to construct eukaryotic recombinant plasmid pEGFP N1 HCV / C + E1. The mouse myeloma cells were transfected with SP2 / 0 and the expression of the green fluorescent fusion protein was observed under a fluorescence microscope. As a result, green fluorescence appeared in the cytoplasm, indicating that the target gene was expressed. And then screened by G418 after a large number of cultured cells for cytotoxicity test target cells, the results show that EGFP reporter gene as a selection marker prepared target cells can fully meet the requirements of cytotoxicity experiments