| 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。 |
| 关键词: 破伤风毒素 C基因 免疫原性 C蛋白 |
| DOI： |
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| Study on cloning,expression and immunogenicity of the fragment C of tetanuss toxin |
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| Abstract: |
| 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 high-expression vector pET32a,and expression of this plasmid in Escherichia coli BL21(DE3) resulted in the production of a fusion protein.The recombinant fragment-Cthioredoxin 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. |
| Key words: tetanus toxin fragment C immunogenicity, protein C |
