摘要
为制备鸭源新城疫病毒M蛋白的多克隆抗体,根据鸭新城疫病毒M基因序列设计1对特异性引物,RT-PCR方法扩增出M基因,克隆入原核表达载体p ET-30a,转化大肠杆菌BL21(DE3),在IPTG的诱导下成功表达重组蛋白,SDS-PAGE结果显示,表达的重组蛋白分子质量约为39 ku。将目的蛋白切胶免疫ICR小鼠,制备重组蛋白的多抗血清,Western blot分析显示,制备的多抗血清能与鸭新城疫病毒感染的SPF鸡胚尿囊液总蛋白发生特异反应。以上结果表明,M基因在大肠杆菌中获得了成功表达,且制备的鼠多抗血清可以用于M蛋白的检测。
In order to express M protein of duck-origin Newcastle disease virus and prepare polyclonal antibodies,according to the genome sequences of duck NDV,one pair of specific primers was designed.The M gene was amplified by RT-PCR,and then was cloned into prokaryotic expression vector p ET-30 a,and the recombinant plasmids were transformed into BL21（ DE3） E. coli. Then the recombinant proteins were successfully expressed following IPTG induction,SDS-PAGE showed that the approximate molecular weight of recombinant expression proteins was 39 ku. The antiserum against the recombinant proteins were produced by immunized ICR mouse with recombinant proteins. Western blot results showed that the antiserum could react specifically with NDV allantoic fluid. These results showed that M gene was successfully expressed in E. coli,and the antiserum could be used for detection of M proteins,and these lay the foundation for the development of detection kit.
出处
《河南农业科学》
CSCD
北大核心
2016年第1期124-126,共3页
Journal of Henan Agricultural Sciences
基金
国家自然科学基金项目(31302096)
江苏省农业支撑项目(BE2013415)
江苏省六大人才高峰项目(NY-009)
