重组白假丝酵母菌Sap2的免疫保护性研究

Study on immunoprotection of recombinant Sap2 of Candida Albicans

目的:构建白假丝酵母菌天冬氨酸蛋白酶2(Sap2)的原核表达载体并诱导表达目的蛋白,进行小鼠试验观察免疫保护效果。方法:PCR克隆白假丝酵母菌Sap2,与pMD19-T-Sap2载体转化至DH5α,对阳性pMD19-T-Sap2质粒和pET32a载体进行双酶切,构建pET32a-Sap2表达载体,转化至BL21(DE3),诱导目的蛋白表达、收集纯化。小鼠行3次免疫接种重组蛋白后,制造系统性白假丝酵母菌感染模型观察小鼠生存期、肾脏病理切片。结果:成功构建pET32a-Sap2原核表达载体,目的蛋白被表达。重组蛋白对小鼠系统性白假丝酵母菌有一定的保护性。结论:接种原核表达的重组白假丝酵母菌Sap2蛋白,能在一定程度上抵抗系统性白假丝酵母菌感染,这为研究Sap2的疫苗奠定基础。

Objective：To construct prokai^cotic expression vector carrying Sap2 of Candida Albicans and express Sap2 protein. Then we aimed to perform the combined immunization on the mouse model by using recombinant Sap2 in order to observe the protective effects on systemic Canddida Albicans infection. Methods： The Sap2 gene of Candida Albicans was amplified by PCR, and the pMD19- T-Sap2 recombinant vector was transformed into DHSc~. The positive PMD19-T- Sap2 and pET32a were digested by double restriction enzymes to construct pET32a-sap2 ,then pET32a-sap2 was transformed into BI21 （DE3） to express the Sap2 protein. The mice were immunizated by inoculating recombinant Sap2 protein for three times. Then the mouse model of Systemic Candida Albicans Infection was established and the immunoprotection was observed. Results： Prokaryotie expression vector pET32a-sap2 was constructed and the Sap2 protein was expressed. Recombination of Sap2 effectively prolonged the mouse survival time, had a good protection to kidney of the mouse. Conclusion： The Sap2 protein of Candida Albicans was obtained by prokaryotie expression system. Sap2 protein had potential to resist Systemic Candida Albicans Infection. This study facilitated the development of new combined Sap2 vaccine.