HBV/HEV融合抗原大肠杆菌表达包涵体的变复性和纯化  

Denaturation,Renaturation and Purification of Inclusion Bodies of HBV/HEV fusion Antigen Expressed In E.coli

作　　者：刘葳[1] 马德章[1] 于航[1] 李冰[1] 于源华[1] 

机构地区：[1]长春理工大学生命科学技术学院,长春130022

出　　处：《长春理工大学学报（自然科学版）》2008年第4期7-11,共5页Journal of Changchun University of Science and Technology(Natural Science Edition)

基　　金：吉林省教育厅资助项目(JYT200508)

摘　　要：利用乙型肝炎病毒adr型的S抗原基因和戊型肝炎病毒China-D型的ORF2编码区的羧基末端中的部分序列构建融合抗原表达载体,在大肠杆菌中表达时,出现了大量包涵体,我们对包涵体进行洗涤,变性,复性,金属亲和层析柱和凝胶过滤纯化后,蛋白的相对含量达98%。采用Western blot对目的蛋白活性进行检测,具有乙肝和戊型肝抗原活性。为提高资源利用效率,降低成本,提高产量,以及进一步研发疫苗用抗原提供了实验基础。When the HBV/HEV fusion antigen expression vector constructed by S gene ofadr subtype of HBV and some sequences located in C-terminal in ORF2 encode region ofchina-D subtype of HEV was expressed in E. coli, a large amount of inclusion bodies emerged.We study on cleaning; denaturation; renaturation of the inclusion bodies and purification through Ni column affinity chromatography and gel filtration. The result indicated that, the relative content of the protein of is 98%, and the protein have all antigen activity of HBV and HEV through Western blot assay.The result of the study can be used to improve the efficiency of resource use, reduce costs, increase output, as well as further development of vaccine antigens.

关 键 词：HBV/HEV融合抗原 包涵体 变性 复性 纯化 

分 类 号：Q789[生物学—分子生物学]