摘要
利用PCR技术扩增获得C型口蹄疫病毒VP1基因,将其克隆到原核表达载体pET-30α(+)上,构建重组表达质粒pET-VP1。质粒pET-VP1转化大肠杆菌BL21(DE3)感受态细胞,用IPTG诱导VP1基因的表达,收集不同时间的菌液进行SDS-PAGE、Western blot分析,结果得到分子量约为33 ku的目的条带,表达产物占菌体总蛋白的35%,且该目的条带能被C型FMDV阳性血清识别,说明VP1基因在大肠杆菌中得到高效表达。融合表达蛋白经镍柱纯化,重组蛋白纯度达90%以上。
Foot-and-mouth disease virus （FMDV） VP1 gene was amplified by RT-PCR. The VP1 fragment was inserted into plasmid pET-30α （＋） to obtain the recombinant plasmid pET-VP1. The pET-VP1 was transformed into Escherichia coli BL-21 （DE3） and induced to ex- press VP1 protein by IPTG. The fusion protein band with a molecular weight of 33 ku was visible on the SDS-PAGE gel. The density scanning showed that the largest amount of the fusion protein was 35% of total bacterial protein. Western blot result showed that the expression product could specifically react with the antiserum to FMDV serotype C. The fusion protein was further purified by Ni-NTA and the purified protein accounted for 90% of total protein.
出处
《畜牧与兽医》
北大核心
2009年第7期4-7,共4页
Animal Husbandry & Veterinary Medicine
基金
国家支撑计划(2006BAD06A14)
关键词
C型口蹄疫病毒
VP1基因
表达
蛋白纯化
foot-and-mouth disease virus （FMDV）
VP1 gene
expression
protein purification
