作　　者： ; ; ; ; ;

机构地区： 华南农业大学兽医学院

出　　处： 《华南农业大学学报》 2003年第3期69-72,共4页

摘　　要： 根据伪狂犬病毒(PRV)NIA 3和Rice株的gE基因序列,设计了1对含有起始密码子和HindⅢ、BamHⅠ酶切位点的引物,以PRV粤A毒株(PRV YA)基因组为模板,利用聚合酶链反应(PCR)扩增获得gE基因片段,将这一片段克隆至PMD18 T载体中,获得重组质粒PMD18 gE;用HindⅢ和BamHⅠ酶切该重组质粒,回收得到含有以上2个酶切位点粘端的gE基因,将此基因片段克隆至相同酶切、回收后的pcDNA3 1(+)真核表达载体中,获得真核表达重组质粒pcDNA3 1 gE,经PCR鉴定、限制性内切酶分析和克隆片段的序列测定、比较,证实了克隆片段的可靠性. Based on the gE nucleotide sequence of PRV strains NIA3 and Rice, a pair of primers were designed. Using the genome DNA of the strain YueA of PRV , which was isolated from an outbreak in Guangdong Province, as template, a segment of gE was amplified by polymerase chain reaction(PCR), and a fragment with the expected size was obtained. The gE gene fragment was cloned into PMD18T vector, and the recombinant plasmid named PMD18gE obtained. The gE fragment was recovered after the plasmid PMD18gE was digested with HindⅢand BamHⅠ, then was cloned into the mammalian expression vector pcDNA31(+). Finally, a recombinant plasmid named pcDNA31gE, which was identified by PCR, restriction enzyme analysis and sequencing ,was obtained.

关 键 词： 猪 伪狂犬病毒粤 株 基因 真核表达 质粒构建 酶切位点 序列测定

领　　域： [农业科学] [农业科学] [农业科学]