破伤风毒素C基因的克隆及重组C蛋白的表达和免疫原性研究  

Study on cloning,expression and immunogenicity of the fragment C of tetanuss toxin

作　　者：蒋会婷[1] 于三科[1] 冯平[1] 江海燕[2] 王希良[2] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《西北农林科技大学学报（自然科学版）》2006年第4期19-22,27,共5页Journal of Northwest A&F University(Natural Science Edition)

摘　　要：对破伤风毒素C基因(TTC)进行了克隆,在大肠杆菌中构建了C基因-硫氧还蛋白融合表达系统并对其进行了重组表达,对表达后蛋白进行了纯化和rTTC的免疫原性初步研究。结果表明,该系统可高水平表达可溶性rTTC,rTTC表达量占可溶性蛋白的28.19%,纯化后纯度为96.92%;所获得的rTTC具有良好的免疫原性,可为开发新一代破伤风亚单位疫苗奠定基础。To clone, express, purify protein and analyse the immunogenicity of the fragment C of tetanus toxin,the fragment C gene of tetanus toxin was amplified from Clostridium tetani genomic DNA by PCR. It was inserted into the hlgh-expresslon vector pET32a ,and expression of this plasmid in Escherichia coli BL21 （DE3） resulted in the production of a fusion protein. The recombinant fragment-C-thioredoxin proteins were purified significantly in one step by affinity chromatography ,and then used to immunize mice and test the antibody titers. The result of SDS-PAGE showed that a specific recombinant protein was expressed,accounting for 28.19% of the soluble protein. Western blot analysis indicated that this product had certain antigenicity,and the final purity was 96.92%. 14 days after the third immunization the anti-tetanus antibody titers of the mice of the group rTTC were detected. The recombinant protein is an immunogenicity antigen of tetanus toxin and thus constructs a basis for the development of vaccine in the future.

关 键 词：破伤风毒素 C基因 免疫原性 C蛋白 

分 类 号：Q784[生物学—分子生物学] Q785