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目的探讨HBcAg显性失活突变体质粒pCDA4-C-GFP的构建及体外表达。方法 选择编码乙肝核心抗原C基因片段及绿色荧光蛋白（green fluorescent protein,GFP）基因片段,利用基因重组技术构建成DNA质粒pCDNA4-C-GFP,并将该质粒转染肝癌细胞株HepG2。结果 通过RT-PCR检测到其RNA的表达,Confocal观察到GFP绿色荧光。结论 HBcAg显性失活突变体质粒pCDNA4-C-GFP的构建成功可以进行对HBV作用的研究。
Objective To investigate the construction and in vitro expression of pCDA4-C-GFP, a dominant negative mutant of HBcAg. Methods The gene encoding the hepatitis C core antigen C gene and green fluorescent protein (GFP) gene were selected and cloned into the DNA plasmid pCDNA4-C-GFP by gene recombination technique. The plasmid was transfected into HepG2 hepatocellular carcinoma cell line. Results The RNA expression was detected by RT-PCR and GFP green fluorescence was observed by Confocal. Conclusion The construction of plasmid pCDNA4-C-GFP with dominant negative mutant of HBcAg can be used to study the effect of HBV.