摘要
[目的]研究鸡传染性支气管炎病毒S1基因的原核表达及反应原性鉴定。[方法]将S1基因克隆到pMD18-T质粒载体上构建pMD18-T-IBV-S1载体;将目的基因插入原核表达载体pET-32a(+)的多克隆位点区,并将构建的重组质粒转化至宿主菌BL21中;IPTG诱导后的蛋白做SDS-PAGE电泳、免疫印迹分析。[结果]S1基因在大肠杆菌中成功地进行了表达,表达的融合蛋白约为66kD,以包涵体形式存在。免疫印迹试验显示,重组蛋白可被IBV多克隆抗体识别。[结论]S1重组蛋白具有反应原性,为进一步研究IBV的新型疫苗奠定了基础。
[Objective] To study the prokaryotic expression and antigenicity identification of S1 gene from avian infectious bronchitis virus（IBV）.[Method] The S1 gene was subcloned into a vector pMD18-T,yielding a recombinant plasmids pMD18-T-IBV-S1,then S1 gene was inserted into the multi-cloning site of a prokaryotic expression vector pET-32a（＋）.The recombinant plasmid was transformed into E.coli BL21.The recombinant protein was induced by IPTG and measured by SDS-PAGE and western-blotting.[Result] The S1 gene was successfully expressed in E.coli BL21,the fusion proteins were about 66 kD in a form of inclusion bodies.Western-blotting test showed that the recombinant proteins can be identified by IBV polyclonal antibody.[Conclusion] The recombinant proteins of S1 gene have reactogenicity,which lays a good foundation to further research on new generation vaccine of IBV.
出处
《安徽农业科学》
CAS
北大核心
2010年第20期10692-10694,共3页
Journal of Anhui Agricultural Sciences
基金
北京市科技新星计划项目(2005B35)资助
关键词
鸡传染性支气管炎病毒
S1基因
原核表达
免疫印迹
Avian infectious bronchitis virus
S1 gene
Prokaryotic expression
Western-blotting
