EB病毒gp125蛋白的酵母表达及在鼻咽癌检测中的应用

发布时间：2018-06-30 17:06

本文选题：鼻咽癌 + gp　；参考：《中山大学学报(医学科学版)》2014年05期

【摘要】：【目的】采用毕赤酵母系统构建表达EB病毒gp125重组蛋白,并探讨其在鼻咽癌血清学诊断中的应用。【方法】以EBV DNA为模版,采用PCR法扩增gp125的编码基因BALF4(2574 bp),与毕赤酵母表达载体pPIC9连接,转化酵母菌GS115,甲醇诱导重组蛋白表达。重组蛋白经SDS-PAGE、免疫印迹法鉴定后,包被聚丙乙烯板,制备ELISA试剂检测鼻咽癌患者和正常人群gp125-IgA抗体。【结果】在毕赤酵母菌中高效地表达了分泌型的gp125重组蛋白,相对分子质量为110 000,免疫印迹证实目的蛋白有免疫原性,gp125蛋白经纯化后作为包被抗原检测鼻咽癌患者和健康对照的敏感度和特异度分别为78.0%和92.0%。【结论】采用毕赤酵母系统高效分泌表达了gp125重组蛋白,初步评估了gp125蛋白在鼻咽癌血清筛选中的诊断价值。
[Abstract]:[objective] to construct and express EBV gp125 recombinant protein by Pichia pastoris, and to explore its application in serological diagnosis of nasopharyngeal carcinoma. [methods] EBV-DNA was used as template. The gp125 encoding gene BALF4 (2574 bp),) was ligated with Pichia pastoris expression vector pPIC9 and transformed into yeast GS115. The recombinant protein was induced by methanol. The recombinant protein was identified by SDS-PAGEand Western blot and coated with polypropyleneplate to prepare Elisa reagent to detect gp125-IgA antibody in patients with nasopharyngeal carcinoma and normal population. [results] the secreted recombinant gp125 protein was expressed efficiently in Pichia pastoris. The relative molecular weight was 110,000. The sensitivity and specificity of immunogenicity gp125 protein was 78.0% and 92.0% in NPC patients and healthy controls, respectively. [conclusion] The recombinant gp125 protein was secreted efficiently by the yeast system. The diagnostic value of gp125 protein in serum screening of nasopharyngeal carcinoma was preliminarily evaluated.
【作者单位】：中山大学附属第三医院检验科;中山大学附属第三医院超声科;
【基金】：广东省科技计划项目(2006B36001010) 教育部第44批留学回国人员科研启动基金[教外司留(2012)940]
【分类号】：R739.63

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