摘要： 根据猪繁殖与呼吸综合症病毒已知序列(ch-1a,AY032626)设计包含完整ORF5序列的1对引物,采用RT-PCR方法扩增PRRSV甘肃LZH株的ORF5基因,将其克隆到杆状病毒载体pFastBacHTA上,经酶切鉴定、PCR鉴定筛选出阳性重组转移载体pFastBacHTA-ORF5,并对阳性质粒进行测序及序列分析 将pFastBacHTA-ORF5转化到DH10Bac感受态细胞中,与Bacmid发生位点特异性转座作用,获得重组转座子rBacmid-ORF5 并成功构建了PRRSV LZH株ORF5基因的杆状病毒载体,为基因工程疫苗的研制奠定了基础。

关键词: 猪繁殖与呼吸综合征病毒, ORF5基因, 克隆, 杆状病毒表达

Abstract: According to the published genome sequences of PRRSV, one pair of spec cprimers were designed toamplify ORF5 gene of L211 strain of GAMS U province by RT-PCR. The ampl}ied fragment was cloned into pFastBacHTAdonor plasmid. The recombinant pFastBacHTA was identified lry digestion with clonuclease enzyme, PCR amplificationand sequencing.Then the purified plasmidwas transformed intDH10Bae-host cells, producing specifc trarsposition withBacmid to yield the recombinant shuttle vehicles, rBacmid-ORF5. The baculovirus cpression vector was successfully corerstrutted, which would be useful for proluction of gene vaccination in future.

Key words: Porcine reproductive and respiratory syndrome virus, ORF5 gene, Cloning, Baculovirus expression

中图分类号: 

• S852.65