绿色荧光蛋白基因与乙肝病毒e抗原基因在家蚕细胞中的表达

Expression of Green Fluorescent Protein Gene and HBsAg Gene in Silk Worm Cells

用绿色荧光蛋白基因（ｇｆｐ）和乙型肝炎病毒（ＨＢＶ）ｅ抗原基因融合，构建了家蚕核型多角体病毒（ＢｍＮＰＶ）重组转移载体ｐＢｍＧＨｅ，使融合基因处于多角体基因（ｐｌｈ）启动子控制之下。共转染后得到的重组病毒ｒＢｍＧＨｅ能在家蚕细胞中表达约５０ｋＤａ的ＧＦＰ／ＨＢＶｅ融合蛋白。被感染的细胞在紫外光下发射出美丽的绿色荧光。用ＥＬＩＳＡ检测，ＨＢｅＡｇ抗原活性滴度达到１１２８００。这种既能发射绿色荧光又具有抗原活性的双功能融合蛋白为研制一种新型的发光免疫诊断试剂开辟了一条新路。

A novel transfer vector pBmGHe with a fusion gene of green fluorescent protein(GFP) and HBeAg under the control of polyhedrin gene promoter was constructed.The recombinant Bombyx mori nuclear polyhedriosis virus(rBmNPV) rBmGHe can express GFP/HBV e fusion protein with MW of～50kDa.The green light is produced from BmN cells when infected with rBmGHe and exited by UV.The antigenicity of HBeAg in fusion protein can be diluted up to 112800 by ELISA analysis.It is probably a good way for a novel diagnosis reagent to HBV because the fusion protein GFP/HBe has double function of green fluorescence and antigenicity.