Taura综合征病毒主要结构蛋白的表达与纯化

Expression and Purification of Major Structure Protein Genesof Taura Syndrome Virus

根据Taura综合征病毒(TSV)基因组,设计特异性引物,从感染病毒组织中提取组织总RNA后扩增,分别将3个主要结构蛋白基因VP1、VP2和VP3克隆到pGEM TEasyVector.与表达载体连接后,导入大肠杆菌中诱导表达,并纯化目的蛋白.诱导表达的融合蛋白分子量分别为54.2×103、43×103和57.1×103,在变性条件下过柱纯化VP1和VP2,一次可以纯化10mg以上纯度较高的蛋白.

According to the genome of taura syndrome virus, the specific primers were designed to amplify the major structure proteins genes (vp1, vp2 and vp3). The amplified products were cloned into the pGEM-T easy vector. In order to construct the recombinant expressions plasmids, the genes of vp1, vp2 and vp3 were cloned to the expression Vectors pPROEXHT-a , pPROEXHT-b and pGEX-KG respectively. After IPTG induction and SDS-PAGE analysis, the fusion proteins of about 54.2×10~3, 43×10~3 and 51.4×10~3 in molecule weight were detected. More than 10mg proteins of vp1 and vp2 were purified under denaturing condition.