从全长cDNA克隆恢复猪瘟病毒方法的研究  

The Study of Recovered Infectious Classical Swine Fever Virus from Full-length cDNA Clone

作　　者：范运峰[1,2] 赵启祖[1] 赵耘[1] 邹兴启[1] 张仲秋[1,2] 王琴[1] 宁宜宝[1] 

机构地区：[1]中国兽医药品监察所,北京100081 [2]中国动物疫病预防控制中心,北京100026

出　　处：《中国畜牧兽医》2009年第6期56-60,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(30571377);国家863计划(2006AA10A204)

摘　　要：以携带猪瘟病毒Thiverval株全长cDNA克隆的pAC/F101/T1-7载体质粒为模板,在体外转录病毒基因组RNA,并转染PK-15和BHK-21细胞,通过传代、RT-PCR、免疫过氧化物酶细胞单层试验鉴定,成功地在两种细胞中拯救出具有感染性的病毒粒子。同时,通过2种细胞转染效率的对比试验,成功建立了利用高转染效率的其它真核细胞作为病毒拯救的过渡细胞,然后再在猪肾细胞系上进行增殖的病毒拯救方式,极大提高了猪瘟病毒拯救的效率。The viral RNA was synthesized in vitro with T7 RNA polymerase using recombinant plasmid pAC/F101/T1-7 as a model, which contained the complete genome of CSFV Thiverval strain, and transfected into PK-15 and BHK-21 cells. Af- ter the cell passage, RT-PCR detection and immunoperoxidase monolayer assay with E2 monoclonal antibody, the results indi- cated that the virus had been rescued successfully from the Thiverval strain full-length cDNA clone in two cells. At the same time, through the comparison of transfected efficiency of PK-15 and BHK-21 cells, the new method of CSFV virus rescue was established that using the high thansfected efficiency cells as transfection and porcine kidney cells as multiplication. This method highly enhanced efficiency of CSFV virus rescue.

关 键 词：猪瘟病毒 疫苗Thiverval株 全长CDNA克隆 病毒拯救 

分 类 号：S852.659.6[农业科学—基础兽医学]