机构地区: 华中农业大学动物医学院
出 处: 《畜牧兽医学报》 2004年第2期192-197,共6页
摘 要: 设计1对引物从含有乙型脑炎病毒NS1基因的质粒pNS1上亚克隆NS1基因,将NS1基因插入到中间转移载体pUSK中,获得重组中间转移质粒pUSK NS1。将pUSK NS1与伪狂犬病毒Ea株TK /gG /LacZ+突变株基因组共转染真核细胞IBRS 2,通过空斑纯化得到了乙型脑炎病毒NS1基因重组伪狂犬病毒株TK /gG /NS1+。经检测,重组病毒能表达具有生物活性的NS1蛋白。该重组病毒可作为猪乙型脑炎和伪狂犬病双价基因工程疫苗用毒株。 One pair of primers was designed to subclone NS1 from pTNS1 into general transfer vector pUSK.A co- transfection experiment was carried out with the purified pUSK- NS1 and the genome of TK^- /gG^- /LacZ^+ mutant of pseudorabies virus Ea strain in IBRS- 2.By plaque purifications and PCR detection,we acquired recombinant virus TK^- /gG^- /NS1^+.Western- blot and ELISA showed that NS1 protein could be expressed in PK- 15 cell infected with TK^- /gG^- /NS1^+.This recombinant virus strain can be used for the study of duel- valence vaccine of JEV and PRV.