水泡性口炎病毒N蛋白原核表达及免疫原性分析

发布时间：2018-04-24 02:43

  本文选题：水泡性口炎病毒(VSV) + N蛋白　； 参考：《中国畜牧兽医》2017年09期

【摘要】：本研究旨在克隆和表达水泡性口炎病毒(vesicular stomatitis virus,VSV)特异性抗原N蛋白,进而纯化并分析其免疫原性。根据GenBank中已发表的VSV基因组N基因序列,分别合成VSV两种不同血清型的N基因,经序列对比分析后,设计合成1对特异性引物,PCR扩增获得约1 300bp的N基因片段,将目的片段亚克隆至pColdⅠ原核表达载体中,经IPTG诱导表达后,采用Ni-NTA树脂亲和层析法纯化重组N蛋白。SDS-PAGE分析表明,N基因在大肠杆菌中得到表达,蛋白大小约为50ku;Western blotting检测结果表明,该重组蛋白与VSV多克隆抗体发生特异性反应。本试验成功构建了VSV-IND和VSV-NJ的原核表达载体,实现了N蛋白在大肠杆菌中的可溶性表达,纯化后的重组蛋白具有良好的免疫原性。
[Abstract]:The aim of this study was to clone and express the specific antigen N protein of vesicular stomatitis virus, and then purify and analyze its immunogenicity. According to the N gene sequence of VSV genome published in GenBank, the N gene of two different serotypes of VSV was synthesized. After sequence comparison and analysis, a pair of specific primers were designed and synthesized to amplify the N gene fragment of about 1 300bp. The target fragment was subcloned into the prokaryotic expression vector of pCold 鈪，

本文编号：1794802