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

doi:10.16431/j.cnki.1671-7236.2017.07.027

Abstract

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.

Key words: infectious bursal disease virus (IBDV); capsid protein; soluble expression; purification

CLC Number:

S858.31

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. Soluble Expression, Purification and Activity Analysis of Capsid Protein of Very Virulent Infectious Bursal Disease Virus[J]. , 2017, 44(7): 2096-2102.

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Soluble Expression, Purification and Activity Analysis of Capsid Protein of Very Virulent Infectious Bursal Disease Virus

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