含双拷贝CSFV E2基因A和D片段原核表达载体的构建及表达

猪瘟是严重危害养猪业的传染病,该病具有高度接触传染性,其流行性广,发病率高,死亡率高,危害极大,被世界动物卫生组织(OIE)列为A类传染病之一。本病的主要临床特征是高热、实质器官出血、淋巴细胞和血小板减少,而怀孕母猪多发生繁殖防碍。每年,猪瘟的发生都为国家造成巨大的经济损失。猪瘟的病原是猪瘟病毒,属于黄病毒科瘟病毒属成员,是单股正链RNA病毒1]。囊膜糖蛋白E2是猪瘟病毒的主要保护性抗原2,3]。E2蛋白可诱导机体产生坚强的中和性免疫保护,具有良好的免疫原性和反应原性4~6] ,这为本病的诊断提供了重要的分子生物学及免疫学…

Construction and Expression of the Prokaryotic Expressing Vector ContainingTandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene

Full E2 gene fragment of Classical swine fever virus (CSFV)shimen strain was amplified by RT-PCR method, then a specific fraction -the A, D antigenic domains- , which encodes protein with good antigenicity and suitable for being expressed in E. coli with high level, was amplified respectively by 2 different pair of primers. Then the 2 amplified fragments were tandemly linked and inserted into prokaryotic expressing vector pET-32a to obtain the plasmid pET-2e. The SDS-PAGE assay showed that although the proteins were present in the form of inclusion body, the linked proteins were expressed from plasmid pET-2e as expected, and can be expressed with high level when induced with IPTG. Western-blotting showed good antigenicity of the target protein.