摘要
应用杆状病毒表达载体成功地表达了汉滩病毒76-118株(HTNV)核壳蛋白。将HTNVS基因插入杆状病毒转染质粒pAcYMIB的多角体基因启动子下游附近,与经Bsu36I酶切线性化的杆状病毒(AcvEPA)DNA共同转染Sf9细胞,经空斑筛选获得了高效表达NP的重组杆状病毒(AcVHanS)。经SDS-PAGE和Westernblot证实,表达产物与HTNV毒粒NP分子量均为50kD左右,紫外扫描显示表达量约占细胞总蛋白量的12.1%,ELISA滴度达1:3200~1:6400。用一批抗NP单克隆抗体和病人血清证实,重组NP与毒粒NP的免疫反应性完全一致。
he recombinant baculovirus, AcVHanS,which contains the S segement of Hantaan Virus was generated by cotransfection of Sf9 cells with the recombinant plasmid (pAcHanS)andBsu36Ⅰlinearized AcVEPA DNA.A high level expression of the HTNV NP was obtained with the baculovirus recombinant.The expressed NP was electrophoretically identified and Wasindistinguishable from the authentic viral NP in molecular weight. Its identity was further con-firmed by Western blot.The expressed NP constituted about 12.1%of total proteins of AcVHanS infected Sf9 cells.The antigen titres by ELISA was 1:3,200-1:6,400.The reactivitieswith HFRS patients’and a panel of MAbs against NP indicated the identity of the ex- pressed NP with the authentic viral NP.
出处
《病毒学报》
CAS
CSCD
北大核心
1995年第2期124-130,共7页
Chinese Journal of Virology
关键词
汉滩病毒
核壳蛋白
杆状病毒
表达
免疫原性
Hantaan virus, Nucleocapsid protein,Baculovirus expression, Immunogenicity