摘要
通过PCR扩增得到产蛋下降综合征病毒(EDSV)纤突蛋白Knob-S区基因,将其克隆至原核表达载体pET-32(a)的多克隆位点构建了原核表达载体pET-32(a)-Knob-S,将其转化至感受态细胞BL21中,经IPTG诱导,SDS-PAGE分析,结果获得了42ku的融合蛋白,用纯化的融合蛋白免疫鸡制备抗血清,通过血凝试验、Western blot和血凝抑制试验分析,表明该融合蛋白不能凝集鸡红细胞,无血凝活性;但可与EDSV阳性血清发生特异反应,并诱导机体产生特异性抗体,具有很好的抗原性。
The Knob-S gene of egg drop syndrome virus（EDSV） was amplified by PCR, and then cloned into the polyclonal sites of pET-32（a） vector. The recombinant prokaryotic expression plasmid was constructed, and named PET-32（a）-Knob-S. The recombinant plasmid was transformed into competent cells BL21 for expression and induced with IPTG. The fusion protein of 42 ku was expressed. Chickens were immunized with the purified fusion protein to prepare polyclonal anti EDSV serum. The hemagglutination test showed that protein could not agglutinate chicken erythrocytes. Western blot test and hemagglutination inhibition test indicated that the expressed protein had good antigenicity.
出处
《动物医学进展》
CSCD
北大核心
2012年第6期41-44,共4页
Progress In Veterinary Medicine
基金
江苏省农业科技自主创新项目(cx(10)450
cx(11)4072
cx(10)216)
