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目的:构建pEGFP/缝隙连接蛋白(connexin,Cx)50真核表达载体,观察其在细胞内的表达和功能情况。方法:经PCR获得Cx50基因片段,重组至有增强绿色荧光蛋白标记的真核表达载体pEGFP-N1中,经脂质体转染至宫颈癌Hela细胞中,通过荧光显微镜检测蛋白表达。结果:转染细胞有增强绿色荧光蛋白及Cx50的表达。结论:融合表达增强绿色荧光蛋白后,不影响缝隙连接蛋白的功能。
Objective: To construct eukaryotic expression vector pEGFP / connexin (Cx) 50 and observe its expression and function in cells. Methods: The Cx50 gene fragment was amplified by PCR and cloned into eukaryotic expression vector pEGFP-N1 with enhanced green fluorescent protein (EGFP). The recombinant plasmid was transfected into cervical cancer Hela cells by lipofectamine 2000. The expression of Cx50 protein was detected by fluorescence microscopy. Results: The transfected cells enhanced the expression of green fluorescent protein and Cx50. Conclusion: The expression of enhanced green fluorescent protein fusion does not affect the function of connexin.