中国动物传染病学报2025，Vol.33Issue(2)：1-10,10.
5种分子伴侣促进牛冠状病毒病毒样颗粒可溶性表达的研究
Five Molecular Chaperones Promote Soluble Expression of Bovine Coronavirus Virus-like Particles
摘要
Abstract
In order to obtain soluble expression of virus-like particles of bovine coronavirus(BCoV)and evaluate their immunogenicity in this study,five molecular chaperones pTf16,pG-Tf2,pG-KJE8,pGro7 and pKJE7 were respectively transferred into E.coli BL21(DE3)to prepare chaperone receptor cells.The recombinant plasmid pET-32a(+)-HBcAg-BCoV S1+S2 carrying the core antigen of hepatitis B virus was transformed into five kinds of chaperone receptor cells.The co-expressing strains were screened by double antibody plates and induced by inducer,and the expression of supernatant was detected by SDS-PAGE to evaluate effective chaperone plasmids.The optimal expression conditions were selected from induction time and inducer concentration.The results showed that the BCoV VLPs were expressed and released into supernatant.The co-expressed recombinant proteins were purified and the formation of VLPs was observed by transmission electron microscopy.Then,mice were immunized with adjuvanted VLPs and the levels of specific IgG and cytokines IFN-γ,TNF-α and IL-4 in serum samples of the immunized mice were detected by ELISA.The results showed that the levels of BCoV IgG and cytokines in mouse serum samples were significantly higher than those of the control group(P＜0.05).Furthermore,the molecular chaperone pG-KJE8 was a better promoter for the soluble expression of BCoV VLPs at 16℃.The results of this study were of great significance for the preparation of economical and efficient BCoV VLP vaccines.关键词
牛冠状病毒病毒样颗粒/可溶性表达/分子伴侣质粒Key words
Bovine coronavirus virus-like particles/soluble expression/molecular chaperone plasmid分类
农业科技引用本文复制引用
郭晓旭,顾文源,赵云环,张帅,范京惠,左玉柱..5种分子伴侣促进牛冠状病毒病毒样颗粒可溶性表达的研究[J].中国动物传染病学报,2025,33(2):1-10,10.基金项目
河北农业大学精准畜牧学科群建设项目(1090064) （1090064）
河北省农业产业技术体系奶牛创新团队项目(HBCT2018120406) （HBCT2018120406）
河北省农业产业技术体系肉牛创新团队项目(HBCT2018130405) （HBCT2018130405）
