摘要
提取猪瘟病毒石门系强毒株的基因组RNA,用RT_PCR扩增其Erns基因的cDNA,将其克隆到原核表达载体pPROEXTMHTc中,然后转化大肠杆菌DH5α,经IPTG诱导,Erns基因获得高效表达。SDS_PAGE分析显示,表达的重组融合蛋白表现分子量约为31ku。Westernblot分析表明,重组蛋白具有良好的免疫原性。重组蛋白可用于制备诊断抗原,用于标记疫苗接种和野毒感染的鉴别诊断。
The cDNA encoding E^(rns) of classical swine fever virus (CSFV) was amplified by RT_PCR from CSFV genomic RNA,and then cloned into an expression vector pPROEX^(TM)HTc.The resulting recombinant plasmid was named pPROC_E^(rns).A His_tagged recombinant protein was expressed with high efficiency in E.coli transformed by pPROC_E^(rns) after induction with IPTG.The fusion protein has an apparent molecular weight of about 31 ku as indicated by SDS_PAGE analysis,and showed specific immunoreactivity with anti_CSFV sera in Western blotting.The recombinant E^(rns) will be used for establishing differential diagnostic assays accompanying marker or DIVA vaccines.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2005年第3期161-163,共3页
Chinese Journal of Preventive Veterinary Medicine
