伪狂犬病病毒Min-A株gE主要抗原表位基因的克隆与鉴定

CLONE AND IDENTIFICATION OF THE PSEUDORABIES VIRUS MIN A STRAIN'S GE MAIN EPITOPES

目的:获得PRVgE主要抗原表位基因,构建PRVgE重组表达载体。方法:根据伪狂犬病病毒Min-A株gE基因序列,设计一对引物,采用PCR方法扩增PRVgE基因主要抗原表位区约642bp的片段,将产物克隆到PGEM-7Z载体中,测序正确后再将其亚克隆到原核表达载体pET-32a中,提取质粒作限制性内切酶分析和序列测定。结果:限制性核酸内切酶酶切鉴定表明,扩增出了PRVgE主要抗原表位基因,测序结果经BLAST分析与GenBank中注册的序列完全一致。结论:成功克隆了PRVgE主要抗原表位基因的原核表达载体。

Objective： To amplify gE gene and construct a fused expression vector of PRV gE gene. Methods： A pair of primers were designed according to the sequence published in the GenBank. A 642 bp fragment encoding gE main epitopes of PRV Min A strain was obtained by PCR and subsequencely cloned into the PGEM- 7Z vector. After it was analyzed with DNA sequencing analysis, the fragment was cloned into PET-32a expression vector. The recombinant plasmid was identified by restriction enzyme analysis and nucleotide sequencing. Resuits： Through restriction endonuclease digestion and DNA sequencing by BLAST analysis completely proved the recombinant plasmid validity. Conclusion： The recombinant expression vector of PRV gE gene was successfully cloned.