【摘要】 目的构建狂犬病病毒G基因和N基因的融合表达载体,导入酿酒酵母表达系统进行诱导表达,为口服基因疫苗的制备打下基础。方法以质粒pVax-G为模板扩增狂犬病毒G和N基因,经连接后与酿酒酵母表达载体pYes2连接,测序鉴定后转入酿酒酵母表达菌株INVScI诱导表达pVax-G/N融合蛋白。结果融合基因pVax-G/N测序结果与预期完全符合,经过半乳糖诱导表达后进行SDS-PAGE电泳和Western-blotting分析,结果表明诱导表达成功。结论成功构建和表达了融合表达载体pYes2-pVax-G/N,为制备以酿酒酵母为运送载体的口服狂犬病病毒疫苗奠定基础。更多还原

【Abstract】 Using pVax-G plasmid as template,G gene and N gene were amplified.A fusion expression vector,named as pVax-G/N,was constructed by linking those two genes together then ligating with Saccharomyces cerevisiae expression vector.Combining with sequence,pVax-G/N was transformed into Saccharomyces cerevisiae expression strain INVScI.Inducing by galactose,the pVax-G/N expressed the correct protein,which proved by SDS-PAGE and Western-blotting analysis.Our experiment provides a basic tool for studying oral rabies virus.更多还原