鸡贫血病毒vp1和vp2基因在毕赤酵母中的表达及产物免疫学特性的初步研究  

Expression of chicken anemia virus vp1 and vp2 gene in Pichia pastoris

作　　者：林欢[1] 高宏雷[1] 王晓艳[2] 高玉龙[1] 宋修庆[1] 李术强[3] 王笑梅[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室／禽传染病研究室,黑龙江哈尔滨150001 [2]中国科学院生物物理研究所,北京100101 [3]哈尔滨工业大学海洋学院,山东威海264209

出　　处：《中国预防兽医学报》2008年第7期495-499,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技支撑计划(2006BAD06A04)

摘　　要：利用特异性引物从pBluemCAV中PCR扩增得到鸡贫血病病毒(CAV)vp1、vp2基因,经EcoRⅠ和NotⅠ双酶切处理,纯化后,克隆至EcoRⅠ和NotⅠ双酶切处理的表达载体pPIC9K中,构建了真核表达质粒pPIC9K-VP1和pPIC9K-VP2。将pPIC9K-VP1和pPIC9K-VP2转化至毕赤酵母SMD1168中,在0.5%甲醇诱导下表达CAV-VP1、CAV-VP2蛋白,运用Westernblot和Dot-ELISA鉴定表达蛋白。结果表明,重组酵母菌株表达出约54ku和24ku的目的蛋白,与针对鸡贫血病毒的单克隆抗体发生特异性反应。将表达产物乳化后免疫6周龄BALB/c小鼠,用ELISA、IFA检测免疫小鼠血清,均检测到抗体。vp1 and vp2 gene from chicken infectious anemia virus （CIAV） were amplified from pBluemCAV by PCR and cloned into pPIC9K vector, respectively. The recombinant plasmid pPIC9K-VP1 and pPIC9K-VP2 were transformed into SMD1168 cells and proteins expressions were induced with methanol. A 54 ku and 24 ku protein were successfully expressed from respective vector. The expression products were shown to react with monoclonal antibodies against CAV in DOT ELISA test. BALB/c mice were immunized with the expressed proteins and specific antibodies were produced and confirmed in ELISA and IFA assays

关 键 词：鸡贫血病毒 VP1基因 VP2基因 真核表达 毕赤酵母 

分 类 号：S852.659[农业科学—基础兽医学]