【摘要】 目的构建增强型绿色荧光蛋白(GFP)与人巨细胞病毒(HCMV)IE1融合蛋白真核细胞表达载体,为IE1基因的表达和功能研究提供基础。方法将质粒pNEB193-IE1和载体pEGFP-C1分别进行双酶切获得目的基因IE1片段和空载体,回收纯化后用T4连接酶将两者连接并转化E.coli JM109,用SalⅠ、BamHⅠ双酶切后琼脂糖电泳鉴定阳性克隆。结果双酶切pEGFP-C1-IE1后,琼脂糖电泳可见到大小约1 476 bp的片段,与IE1预期片段大小一致,即IE1亚克隆入pEGFP-C1。结论成功地构建了HCMV IE1-GFP融合蛋白真核表达载体pEGFP-C1-IE1。更多还原

【Abstract】 Objective To construct the eukaryotic expression vector of enhanced green fluorescent protein and HCMV IE1 fusion protein for exploring the expression and function of IE1 gene.Methods HCMV IE1 gene fragment obtained from pNEB193-IE1 by enzyme digestion was inserted into expression vector pEGFP-C1,then transformed into Escherichia coli JM109,a recombinant pEGFP-C1-IE1 was constructed and confirmed by SalⅠ/BamHⅠdigestion.Results The target gene IE1 fragment about 1476bp was obtained and IE1 was successfully subcloned into pEGFP-C1 vector.Conclusion The recombinant eukaryotic expression vector pEGFP-C1-IE1 was successfully constructed.更多还原