伪狂犬病病毒上海株EP0基因的序列分析及其表达载体的构建

以伪狂犬病病毒上海株 (PRV SH)细胞感染物为模板 ,PCR扩增出 1 2 3kb的EP0基因完整编码区片段 ,将该基因片段克隆到pGEM T easy中 ,经过双脱氧末端终止法序列测定 ,并同国内的PRV Ea株进行同源比较 ,发现PRV SH株EP0基因存在多处突变和一处插入。进一步将该片段插入到原核表达载体 pET32a (+)的His Tag下游 ,构建了的原核表达质粒 pETEP0 ,为今后深入研究该基因的表达及其功能奠定了基础。

Sequencing analysis of EP0 gene of pseudorabies virus SH strain and construction of its expression vector

The EP0 ORF of pseudorabies virus Shanghai strain was obtained by polymerase chain reaction (PCR) using the cells infected with PRV-SH as template, subsequently cloned into pGEM-T-easy vector. The nucleotide was compared with the corresponding sequence of PRV-Ea strain. There were several mutant and a insertion in EP0 gene of PRV-SH strain. Then, the EP0 gene was cloned into pET32a(+), downstream of His-Tag, and the pETEP0 expression vector was constructed, which established a base for further study of expressing and function of the EP0 gene.