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目的表达并纯化粉尘螨2类变应原Der f 2蛋白,检测其变应原性。方法用1mmol/L的IPTG诱导含重组质粒pET30a-Der f2的大肠埃希菌BL21(DE3),SDS-PAGE电泳检测并纯化目的蛋白,ELISA检测其与尘螨哮喘患者血清IgE的结合活性。结果 SDS-PAGE电泳检测重组质粒pET30a-Der f 2表达产物,其分子质量单位为15ku,与预期大小相符。ELISA检测纯化的Der f 2蛋白可与尘螨哮喘患者血清IgE反应。结论成功获得并纯化了Der f2重组蛋白,该蛋白具有变应原性,为进行尘螨过敏性疾病的变应原特异性免疫治疗的动物实验奠定了物质基础。
Objective To express and purify Der f 2 protein, a type 2 allergen of Dust mite, and test its allergenicity. Methods Escherichia coli BL21 (DE3) containing recombinant plasmid pET30a-Der f2 was induced by 1 mmol / L IPTG. The recombinant protein was detected by SDS-PAGE electrophoresis and purified. The binding activity of IgE to serum IgE in dust mite asthma patients was detected by ELISA. Results The recombinant plasmid pET30a-Der f 2 was detected by SDS-PAGE. The molecular mass unit was 15ku, which was consistent with the expected size. The purified Der f 2 protein was detected by ELISA in serum IgE of dust mite asthma patients. Conclusion The Der f2 recombinant protein was successfully obtained and purified. The protein has the allergenicity, which lays the material foundation for the animal experiment of allergen-specific immunotherapy for dust mite allergic diseases.