利用SUMO表达系统高效可溶性表达鸡传染性法氏囊病病毒VP3基因

Use of the SUMO expression system to efficiently express the soluble VP3 protein of infectious bursal disease virus

为评价SUMO原核表达系统(pHisSUMO Express)对病毒基因的可溶性表达,本研究从人工接种发病的鸡传染性法氏囊病(IBD)的病料组织样品中提取总RNA,通过RT-PCR扩增IBD病毒(IBDV)VP3基因,并将其克隆于pHisSUMO中构建了重组表达质粒pHisSUMO-VP3,转化大肠杆菌Rosetta(DE3)PlysS,经IPTG诱导,得到可溶性表达的融合蛋白SUMO-VP3。结果表明,该融合蛋白表达量占细菌总蛋白35%,经HisTrapTMFF crude column层析柱纯化后的SUMO-VP3蛋白可被SUMO蛋白酶Ⅰ有效切割,获得无标签的VP3蛋白,经western blot鉴定表明该VP3蛋白具有良好的抗原性。本研究表明pHisSUMO Express表达系统是高效可溶表达外源蛋白的有效工具,所表达的病毒蛋白具有良好的抗原性,为病原诊断抗原的研究和制备提供有效表达系统。

Efficient expression of the soluble viral proteins is important for virology studies.The aim of this study is to utilize the pHisSUMO expression system for efficient expression of soluble VP3 gene of infectious bursal disease virus（IBDV）.The total RNA of IBDV was used as template to clone the VP3 gene by RT-PCR technology.The VP3 gene was cloned into pHisSUMO expression vector to construct the recombination expression vector pHisSUMO-VP3.High yield and soluble VP3 protein are expressed by pHisSUMO system.The fusion proteins could be efficiently cleaved by SUMO protease-Ⅰ to obtain pure and de-tagged recombinant proteins.This system provides an effective method for virus protein soluble expression de-tagged.