摘要
Abstract
[Objective]This study aimed to achieve the large-scale expression and purification of p26 protein of equine infectious anemia virus(EIAV),so as to address the limitations of conventional methods such as low expression efficiency and high purification costs.[Methods]A prokaryotic expression system for the p26 protein was constructed using recombinant Escherichia coli.The fermentation process utilizing synchronized regulation of multiple parameters was optimized,including induction temperature,pH,IPTG concentration,and induction time,and the expressed protein was purified by nickel affinity chromatography.[Results]The optimal induction conditions were determined as follows:temperature 37℃,pH 7.4,addition of 0.5 mM IPTG at 3.5 h of cultivation,followed by 5 h of induction,under which the expression level of p26 protein reached its maximum.According to SDS PAGE analysis,the molecular mass of the purified protein was consistent with the expected value,meanwhile,results of Western blot analysis demonstrated specific reactivity of the recombinant p26 protein with EIA positive serum,indicating its strong immunoreactivity.[Conclusion]The large-scale expression and purification process for EIAV p26 protein had been successfully established,providing reliable raw material support for EIA serological detection techniques,which was of important application value for improving the monitoring,prevention and control of EIA.关键词
马传染性贫血病毒/p26蛋白/原核表达/蛋白纯化Key words
equine infectious anemia virus/p26 protein/prokaryotic expression/protein purification分类
农业科技
