›› 2007, Vol. 34 ›› Issue (6): 109-111.

• 疾病防治 • 上一篇    下一篇

猪伪狂犬病病毒SA株的分离鉴定及其gE基因缺失株转移载体的构建

管宇1，2，沈志强2，曲光刚2,吕素芳2,任艳玲2,李书光2,王玉炯1
   

  1. 1.宁夏大学生命科学学院，银川 750021;2.山东省滨州畜牧兽医研究院，滨洲 256600
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-06-20 发布日期:2007-06-20

Isolation and Identitication of PRV SA Strain and Construction of gE-delection Transfer Vector

GUAN Yu1,2,SHEN Zhiqiang2,QU Guanggang2,LV Sufang2, REN Yanling2,LI Shuguang2, WANG Yujiong1
   

  1. 1.Ningxia University Life Sciences College, Yinchuan 750021,China;2.Shandong Binzhou Animal Science and Veterinary Medichine Academy, Binzhou 256600,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-06-20 Published:2007-06-20

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pBluescript(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。

关键词: 猪伪狂犬病病毒; 分离鉴定; gE基因; 转移载体

Abstract: 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-delection transfer vector had been successfully constructed.

Key words: PRV; isolation and identitication; gE; transfer vector

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