摘要
质粒pS65T含有T7启动子驱动的绿色荧光蛋白突变型gfpS65T基因，经修饰后，在其中插入乙型肝炎病毒e抗原（HBeAg）基因，使两基因同框成为融合基因。在大肠杆菌BL21中由于T7启动子的控制，高效表达了具有双功能（抗原性和发光性）的融合蛋白（GFP－HBeAg）。融合蛋白MW为52kDa，N端有6个组氨酸残基，因而用Ni金属螯合层析柱对融合蛋白进行了分离纯化，利用诊断HBV的ELISA试剂盒检测了融合蛋白的抗原性，在荧光显微镜下观察到了融合蛋白的绿色荧光。并探讨了用其组装成新型免疫诊断试剂的可能性。
The bi - function fusion protein of hepatitis B virus e antigen and mutant green fluorescent protein was expressed in E. coli BL21 strain. The fusion protein was separated and its anti-genecity was determined by ELISA kit for HBe, meanwhile the intensity of its luminescence could be observed. The approach in which the antigen was tagged by equal molecular GFP allows using the bi-function fusion protein to establish a new method for immunogical diagnosis.
出处
《中国病毒学》
CSCD
1997年第4期336-341,共6页
Virologica Sinica
基金
世界银行贷款项目
关键词
乙型肝炎病毒
e抗原基因
绿色荧光蛋白
基因
HBeAg gene, Green fluorescent protein gene, T7 phage promotor, Bi-function protein
