白念珠菌磷酸甘油酸激酶1的原核表达及免疫原性分析

发布时间：2017-12-26 23:32

本文关键词：白念珠菌磷酸甘油酸激酶1的原核表达及免疫原性分析　出处：《细胞与分子免疫学杂志》2013年10期 　论文类型：期刊论文

【摘要】：目的构建白念珠菌基因磷酸甘油酸激酶1(PGK-1)基因的原核表达质粒并诱导其表达,对其免疫原性进行分析。方法 PCR法获取白念珠菌SC5314的PGK-1基因,插入原核表达载体pET30a中,以重组质粒转染感受态细胞E.coli BL21,经IPTG诱导表达后,SDS-PAGE检测蛋白的表达情况,并用Western blot法验证。用纯化获取的重组蛋白免疫BALB/c小鼠,ELISA对蛋白的免疫原性进行评价。结果成功构建原核表达质粒pET-30a-pgk-1,诱导后表达的重组蛋白相对分子质量(M r)约为54 810。ELISA检测抗体效价约为1∶1 024。结论成功获得白念珠菌PGK-1重组蛋白,该蛋白具有良好的免疫原性。
[Abstract]:Objective to construct the Candida albicans gene phosphoglycerate kinase 1 (PGK-1) prokaryotic expression plasmid gene and its immunogenicity analysis. Methods the PGK-1 gene of Candida albicans SC5314 was obtained by PCR. The PGK-1 gene was inserted into prokaryotic expression vector pET30a. The recombinant plasmid was transfected into the competent cells E.coli BL21, after IPTG induced expression, SDS-PAGE was used to detect the expression of protein, and verified by Western blot method. The recombinant protein obtained by purification was used to immunize BALB/c mice, and ELISA was used to evaluate the immunogenicity of the protein. Results the prokaryotic expression plasmid pET-30a-pgk-1 was successfully constructed, and the relative molecular mass of recombinant protein (M R) expressed after induction was about 54810. The titer of ELISA was about 1: 1024. Conclusion the recombinant protein of Candida albicans PGK-1 has been successfully obtained. The protein has good immunogenicity.
【作者单位】：解放军白求恩国际和平医院检验实验科;
【基金】：河北省自然科学基金(C2010001882)
【分类号】：R392-33
【正文快照】： 念珠菌是真菌感染最常见的病原体,可引起皮肤、黏膜感染,也可侵入深部器官导致危及生命的重症侵袭性感染。侵袭性念珠菌病(invasive candidia-sis,IC)是指由念珠菌属病原菌引起的系统性真菌感染,其中念珠菌血流感染(bloodstream infection,BSI)占医院获得BSI中的第4位,在重症

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