摘要
利用PCR方法获得 4次重复的猪瘟病毒E2蛋白中和性抗原表位基因 ,将其克隆到 pGEM 5ZF(+)载体 ,测序正确后亚克隆到pGEX 3X载体构建得到重组质粒pGEX 3X 4P。重组质粒在大肠杆菌中诱导表达了含 4重复抗原表位的融合蛋白。该蛋白经纯化后 ,利用间接ELISA检测其与血清的反应性 ,结果表明 ,纯化的融合蛋白与兔抗CSFVE2血清有很强的反应性 ,与兔抗BVDVE2血清不反应 。
The peptide TAVSPTTLR is an antigenic epitope mapped recently on the E2 glycoprotein of classical swine fever virus. Based on its genetic sequence a gene fragment encoding repeatedly this epitope in quadruple form wasconstructed by using PCR and cloned into pGEM 5ZF followed by subcloning into expression vector pGEX 3X, resulting in the construction of pGEX 3X 4P. After transformation of E.coli JM109 with pGEX 3X 4P a 4 repeated epitope peptide of CSFV E2 was expressed in fusion with GST. This fused protein, refered to as GST 4P, was prepared by passing the culture product of transformed JM109 through Microspin GST Purification Module, then characterized for its reactivity respectively with some antisera in ELISA. The results showed that this quadruple epitope peptide could react well with rabbit anti CSFV E2 serum and pig anti CSFV serum, but not with rabbit anti BVDV E2 antiserum. This result indicated that this epitope is CSFV specific and the quadruple epitope constructed in the study was likely to be able to differentiate the infection of CSFV from BVDV in antibody ELISA.
出处
《高技术通讯》
EI
CAS
CSCD
2003年第10期41-45,共5页
Chinese High Technology Letters
基金
973规划 (G19990 1190 3)
国家自然科学基金 (398932 90 1)资助项目
