HBcAg颗粒纯化和鉴定和其透射电镜观察.PDFVIP

维普资讯 第 19卷 2期 中 国 病 毒 学 19 (2)： l0l～ l04 2004年4月 VIROLOGICA SINICA April 2004 酿酒酵母表达的rHBcAg ． 颗粒 的纯化和鉴定及其透射 电镜观察 陈 恒 2一，梁婉琪 ，张大兵3$$ (1．上海交通大学生命科学技术学院，上海 200030；2．上海大学生命科学学院，上海 200436 3．f海市农业科学研究院生物技术中心，上海 201106) PurificationandObservationoftheRecombinantHepatitisB VirusCore Antigen(rHBcAg)ParticlesProducedintheYeast CHEN—Heng ，2’。LIANGWan— qi。ZHANG Da—Bing。 ． lSchooloiL@ ScienceandBiotechnology,ShanghaiJiaotongUniversity,Shanghai200030China；2．SchoolofLife Science，ShanghaiUniversit3．Shanghai200436，China；3 ． BiologicalTechnologyCenterofShanghaiAgricultureScience Institute，Shanghai201106，Chinaj Abstract：HepatitisB viruscoreantigenproteingene (C gene)wasexpressed inSaccharomyces cerevisiaeandtheproducts(rHBcAgparticles)werepurifiedrfom crudelysateoftheyeastbythree steps：SephroseCL一4B chromatography，sucrosestep—gradientultracentrifugation，andCsC1一isopycnic ultracentrifugation．rHBcAg wassynthesized in yeastcellsasaparticleconsisting ofpolypeptides whichhaveamolecularweightof21．5kDa (p21．5)．ELISA andCsC1-isopycnicultracentrifugation purificationproducts(rHBcAgparticles)ineachrfactionindicatedthattheparticledensitiesofthe rfactionwithhigherrHBcAgantigenicitymainlydistributedatthedensitiesof 1．27g ·mL— and 1．40 g。mL～，respectively．ObservationofthepurifiedproductsrHBcAgparticles)byTEM indicatedthat rHBcAgpeptidescouldself-assembleintotwokind