新Ⅰ型鸭肝炎病毒VP1和3D蛋白的原核表达及鉴定  

Expression and identification of VP1-/3D-encoding genes of new duck hepatitis type 1 virus

作　　者：孟凡依[1,2] 殷秀辰[1] 李刚[1] 陈玉环[1] 张云[1] 刘明[1] 冯新畅[2] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所动物细菌病研究室,黑龙江哈尔滨150001 [2]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2012年第2期149-151,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：现代农业产业技术水禽体系岗位科学家专项(CARS-43-10)

摘　　要：为表达新I型亚肝炎病毒(DHV-1C)的VP1和3D重组蛋白,本研究根据GenBank中的DHV-1C N株全基因序列,设计合成两对引物扩增其VP1和3D基因,构建重组表达载体并进行诱导表达。SDS-PAGE和western blot分析表明,VP1和3D重组蛋白分别在诱导3 h和4 h后表达量达到峰值,其大小分别为37.68 ku和65.80 ku,并且能够与抗DHV-1C阳性血清产生特异性免疫反应。To express the VP1-/3D protein of new duck hepatitis type 1 virus （DHV-IC）, the VP1-/3D-encoding genes were amplified by PCR with primers designed according to DHV-1C N strain and cloned into expression vector pET-30a, respectively. The recombinant VP1 and 3D were highly expressed for 3 hours and 4 hours after IPTG induction, respectively. SDS-PAGE analysis showed that the VP1 and 3D products were 37 ku and 57 ku, respectively. Western blot demonstrated that VP1 and 3Dproteins reacted specifically with duck anti DHV-1C serum. The expression of VP1 and 3D protein in bacterial could be useful in the development of diagnostic kit and subunit vaccine, or the study of pathological mechanism of DHV-1C.

关 键 词：新I型鸭肝炎病毒 DHV-1C 3D VP1 克隆表达 

分 类 号：S852.65[农业科学—基础兽医学]