摘要
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.
基金
江苏省自然科学基金
关键词
GFP
BMNPV
乙型肝炎病毒
e抗原基因
融合蛋白
Bombyx morinuclear polyhedrosis virus\ Green fluorescent protein gene\ Fusion protein