【摘要】 以伪狂犬病毒Ea株BamH I 5′片段为模板,PCR扩增出约0.9 kbgK基因完整编码区片段,将该基因克隆到pBluescriptIISK+中,构建重组载体pSKgK。进一步将该片段插入真核表达载体pEGFP-C1的增强绿荧光蛋白EGFP下游,构建真核表达载体pEGFPgK,脂质体法转染BHK-21细胞,G418筛选至正常细胞完全死亡,在倒置荧光显微镜下观察,pEG-FPgK转染的细胞在细胞膜和细胞浆中发出很强的荧光,而正常细胞不发荧光。证明gK基因在BHK-21细胞上获得了表达,表达在细胞膜和细胞浆中。更多还原

【Abstract】 The fragment containing gK gene coding domains was amplified by polymerase chain reaction(PCR) using BamH I 5′ fragment of PRV Ea strain as template.The fragment was cloned into pBluescriptIISK+,resulting recombination plasmid pSKgK.Then the gK gene was cloned into eukaryotic expression vector pEGFP-C1,following the EGFP(Enhanced Green Fluorescent Protein).The recombinant plasmid pEGFPgK were transfected into BHK-21 cell with lipofectin and selected under G418,until the normal BHK-21 cell die out.Expression protein was detected under inverted fluorescence microscope.The cells transfected pEGFPgk excited strong fluorescence in cytomembrane and cytoplasm,The results showed that gK gene was expressed in the cytomembrane and cytoplasm of BHK-21 cell.更多还原