文献类型：期刊文献

中文题名：SUMO融合系统高效表达可溶性IBDV NB株VP2 S-P域蛋白

英文题名：The SUMO Fusion System Efficiently Expresses the Soluble IBDV NB Strain, VP2 S-P Domain Protein

作者：朱艳平[1];邢瑞林[1,2];李润芷[1,3];何勇[1];申一兰[1,4];王玲玲[1,5];闫占鹏[1,2];任鹏举[1,2];徐若楠[1,2];岳锋[1];李鹏[1];王选年[1,2]

第一作者：朱艳平

机构：[1]新乡学院生命科学技术学院,新乡453003;[2]郑州大学生命科学学院,郑州450000;[3]河南科技学院动物科学学院,河南新乡453003;[4]河南师范大学生命科学技术学院,新乡453003;[5]河南科技大学动物科技学院,洛阳471000

第一机构：新乡学院生命科学技术学院

年份：2019

卷号：38

期号：9

起止页码：3929-3934

中文期刊名：基因组学与应用生物学

外文期刊名：Genomics and Applied Biology

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：国家自然基金(3167131497);新乡学院青年骨干教师项目(GGJS2016-06);新乡学院科技创新团队项目(XXUTD20170106; XXUTD20170107);新乡市科技创新平台项目(CP1311)共同资助

语种：中文

中文关键词：鸡,传染性法氏囊病病毒;VP2-S-P;SUMO

外文关键词：Chicken;Infectious bursal disease virus;VP2-S-P;SUMO

摘要：鸡传染性法氏囊病病毒(infectious bursal disease virus, IBDV)导致机体出现免疫抑制。VP2 是IBDV主要保护性抗原表位的结构蛋白,Shell 域和Projection 域位于IBDV VP2 蛋白的胞外区和跨膜区,包含VP2主要的抗原位点。前期研究在Shell 域筛选到能够与IBDV 多抗结合的多肽。因此,本研究利用PCR 扩增Shell 域和Projection 域的基因,添加小泛素相关修饰物(small ubiquitin-related modifier, SUMO)作为标签,构建SUMO系统高效表达系统,诱导表达获得VP2 Shell 域和Projection 域的可溶性融合蛋白,命名为SUMOVP2-S-P。SDS-PAGE和Western blotting鉴定重组蛋白。SDS-PAGE结果显示,出现与预期蛋白大小相符合的可疑条带;Western blotting鉴定结果显示,SUMO-VP2-S-P重组蛋白能够与His 单抗特异性结合。初步结果表明,SUMO-VP2-S-P重组蛋白为我们需要的目的蛋白,为后期蛋白纯化建立IBDV抗体快速检测方法提供材料基础。
Chicken infected with infectious bursal disease Virus (IBDV) could cause immune suppression in the body. VP2 is the structural protein containing the main protective epitopes of IBDV. The Shell domain and the Projection domain are located in the extracellular and transmembrane regions of IBDV VP2 protein, containing the main antigenic sites of VP2. In our previous research, Peptide in the Shell domain could been screened multiple resistance binding to IBDV polypeptide. Therefore, the gene of Shell domain and Projection domain was amplified by PCR, where small Ubiquitin-related Modifier (SUMO) was added as a tag to construct a highly efficient expression system. The soluble fusion protein of VP2 Shell domain and Projection domain induced by IPTG was named as SUMO-VP2-S-P, which was identified with SDS-PAGE and Western blotting. SDS-PAGE results showed that suspicious bands with the expected protein size appeared. Western blotting identification indicated that SUMO-VP2-S-P recombinant protein could be combined with His-tag monoclonal antibody. The preliminary results showed that the SUMO-VP2-S-P recombinant protein was the target protein which we need. It could provide the material basis for the establishment of a fast detection method for IBDV antibody in late protein purification.