家蚕核多角体病毒载体稳定高效表达ＨＢeＡg条件及其初步应用的研究

利用已构建的含有HBeAg基因的重组家蚕核多角体病毒rBmHBe，摸索了HBeAg在家蚕细胞中表达的最佳条件组合，细胞接种密度（3.2～6.4）×10

Study of the High-Production and the Primary Application of the HB eAg Expressed with BmNPV System

The recombinant BmNPV(rBmHBe) was used to study the optimalconditions for expression of HBeAg gene in BmN cell. The results demonstrated that the optimal cell density, time of infection and MOI were（3.2～6.4）×105 cell／flask,48～ 60 h,and 0.04～0.4PFU／cell respectively.Then the HBeAg in the supernatant of BmN cell culture was separated and purified by precipitation with (NH4)2SO4,gel filtration and anion exchange chromatography successively, and the product appeared as a single band on SDS-PAGE.Finally,the purified HBeAg was used to detect the antiHBe by double antigen sandwich ELISA.The sensitivity of the method was super to that of antiHBe kit from market.In addition,the cross reactivity of HBeAg from BmN cell with antiHBc as 100 times as low as that of HBe Ag from E.coli. Meanwhile the antigenicity of HBeAg from BmN cell was 100 times as high as that of HBeAg from E.coli.