摘要
目的 探索原核可溶性目的蛋白表达系统对狂犬病病毒糖蛋白膜外区（Ex-GP）进行高效制备及纯化的方法.方法 构建含有融合标签的狂犬病病毒糖蛋白膜外区表达质粒,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,以获得高效表达的可溶性目的蛋白.结果 成功建立应用原核表达系统高效制备重组可溶性狂犬病病毒糖蛋白膜外区蛋白的方法,获得的目的蛋白具有ELISA抗原活性,推测其构象与天然狂犬病病毒糖蛋白膜外区具有相似性.结论 本研究建立的目的蛋白高效制备方法,操作简便,表达量高,目的蛋白具有抗原性活性,适合目的蛋白规模制备,为深入研究狂犬病病毒糖蛋白生物学功能,解析糖蛋白结构,揭示狂犬病病毒致病机制及开展狂犬病诊断试剂的研发提供了思路.
Objective To express and purify the glycoprotein extracellular domain （Ex-GP） of Rabies virus strain CTN in soluble form with high efficiency.Methods A recombinant expression plasmid containing the gene encoding the Ex-GP was constructed.Various expression conditions were screened to obtain an optimum prokaryotic expression system for Ex-GP in soluble form.The expressed target protein was purified using affinity chromatography and gel filtration chromatography.Results The target protein Ex-GP with high antigenicity was efficiently expressed in soluble form by using the recombinant PBCX expression system and effectively purified by using affinity and gel filtration chromatography.Conclusion The soluble form of Ex-GP is successfully expressed and purified in a simple and convenient way.This study paves the way for further researches on the biological functions of rabies virus glycoprotein,the pathogenic mechanism of rabies and the development of diagnostic reagent and vaccines for rabies virus.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2017年第6期438-442,共5页
Chinese Journal of Microbiology and Immunology
关键词
狂犬病病毒
糖蛋白膜外区
可溶性表达
Rabies virus
Glycoprotein extracellular domain
Soluble protein expression
