中国畜牧兽医2017，Vol.44Issue(7)：2096-2102,7.DOI:10.16431/j.cnki.1671-7236.2017.07.027
传染性法氏囊病病毒超强毒株衣壳蛋白的可溶性表达、纯化与活性分析
Soluble Expression, Purification and Activity Analysis of Capsid Protein of Very Virulent Infectious Bursal Disease Virus
摘要
Abstract
To obtain the capsid VP2 with high quality, VP2 gene of very virulent infectious bursal disease virus (vvIBDV) Gx was cloned and inserted into pCold-Ⅰ and the prokaryotic expression plasmid pCold-Ⅰ-GxVP2 was constructed.In engineering bacteria Transetta(DE3), the induction conditions of protein VP2 expression were optimized.With affinity chromatography and gel filtration, protein VP2 was purified.With the monoclonal antibody directed Western blotting, protein VP2 was identified.Using SPF chicken, immunocompetence of VP2 was evaluated.The results showed that the dissoluble protein VP2 was expressed successfully in Transetta(DE3) in cold-shock conditions;Protein VP2 was purified and the concentration was 542 μg/mL;The purified protein VP2 not only reacted with the monoclonal antibody against protein VP2, but also induced specific immune response in immunized chickens.In general, with 15 ℃ of cold-shock condition, 120 r/min of shaking culture, 1 mmol/L of IPTG,inducting for 24 h, soluble capsid VP2 of IBDV with immunocompetence was successfully expressed and purified.The preparation of highly purified, soluble capsid protein with functional activity laid the foundation for further researches on the pathogenic mechanism.关键词
传染性法氏囊病病毒/衣壳蛋白/可溶性表达/纯化Key words
infectious bursal disease virus (IBDV)/capsid protein/soluble expression/purification分类
农业科技引用本文复制引用
高祥,王笑梅,祁小乐,李辉,张礼洲,卢珍,王永强,高立,吴甜甜,高玉龙,刘长军..传染性法氏囊病病毒超强毒株衣壳蛋白的可溶性表达、纯化与活性分析[J].中国畜牧兽医,2017,44(7):2096-2102,7.基金项目
国家自然科学基金重点项目(31430087) （31430087）
哈尔滨市应用技术研究与开发项目(2014AB3AN058) （2014AB3AN058）
现代农业产业技术体系建设专项基金(CARS-42-G07) （CARS-42-G07）
哈尔滨市科技创新人才项目(2014RFQYJ129) （2014RFQYJ129）
