Soluble Expression,Purification and Activity Analysis of Capsid Protein of Very Virulent Infectious Bursal Disease Virus |
| |
| Authors: | GAO Xiang LI Hui ZHANG Li-zhou LU Zhen WANG Yong-qiang GAO Li WU Tian-tian GAO Yu-long LIU Chang-jun WANG Xiao-mei QI Xiao-le |
| |
| Institution: | 1. Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150069, China;
2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China |
| |
| 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. |
| |
| Keywords: | infectious bursal disease virus (IBDV) capsid protein soluble expression purification |
|
| 点击此处可从《中国畜牧兽医》浏览原始摘要信息 |
| 点击此处可从《中国畜牧兽医》下载免费的PDF全文 |
|
