论文部分内容阅读
目的通过原核表达系统对狂犬病病毒CTN株核蛋白(NP)进行高效的可溶性表达及纯化。方法构建含有融合标签的狂犬病病毒CTN株核蛋白表达质粒,筛选可溶性表达条件,采用亲和层析和凝胶过滤层析对目的蛋白进行纯化,切除目的蛋白的融合标签,最终获得高效表达的可溶性目的蛋白。结果成功建立应用PBCX原核表达系统高效表达重组可溶性狂犬病病毒CTN株核蛋白rb-NP的表达及纯化工艺,获得高纯度目的蛋白。结论本研究建立的可溶性表达狂犬病病毒核蛋白的方法,操作简便,表达量高,适合目的蛋白的规模制备,为深入研究狂犬病病毒核蛋白生物学功能、开展狂犬病诊断试剂研究及新型疫苗的研发提供了新的技术手段。
Objective To express efficiently and purify the nucleocapsid protein (NP) of rabies virus CTN strain by prokaryotic expression system. Methods Rabbit virus CTN strain containing fusion protein was constructed, and the soluble expression conditions were screened. The target protein was purified by affinity chromatography and gel filtration chromatography, and the fusion protein was excised and the highly expressed soluble protein was obtained The target protein. Results The prokaryotic expression system of PBCX was successfully constructed to express rb-NP protein of recombinant soluble lyssavirus CTN strain and its purification technology was successfully established. Conclusions The method of soluble expression of rabies virus nucleoprotein established in this study is simple and convenient to operate and has high expression level and is suitable for the scale preparation of target protein. In order to further study the biological function of rabies virus nucleoprotein, research on rabies diagnostic reagent and development of novel vaccine A new technical means.