摘要
从某疑似猪伪狂犬病发病仔猪提取脑组织病料。病料处理后,经vero细胞连续分离培养,盲传7代后出现了典型细胞病变。PCR检测证实该分离病毒为猪伪狂犬病病毒;利用多种传代细胞系对该病毒的培养条件进行了优化,该病毒的最佳培养宿主为BHK-21细胞,且在该细胞的TCID50为10-9.25/0.1 ml。将含有YFP报告基因表达盒PM片段连接到pBlue-script(M13-)载体后,分别克隆猪伪狂犬病病毒gE基因同源重组臂PL、PR片段,并将其连接到上述重组载体上。经鉴定,成功构建了猪伪狂犬病病毒gE基因缺失株转移载体。
The brain Sample of swine that had been doubtfully infected Aujeszky＇s disease was obtained . The sample was treated and inoculated into Vero and the cell indicated CPE after 7 generations. PCR analysis showed the virus was PRV. The virus was inoculated into other cells and the BHK-21 cell was the optimization for the virus with TCID50 10^-9.25/0.1 ml . The PM gene fragment that contains YFP express frame,and the PL and PR fragment used for recombination that were cloned from PRV gE gene,were sequentially inserted into pBluescript（M13-） vector. The results of identitication showed that the gE-delec; tion transfer vector had been successfully constructed.
出处
《中国畜牧兽医》
CAS
2007年第6期109-111,共3页
China Animal Husbandry & Veterinary Medicine
