中国病理生理杂志2013，Vol.29Issue(6)：1147-1152,6.DOI:10.3969/j.issn.1000-4718.2013.06.033
原核表达和制备EB病毒GST/BFRF3融合蛋白用于鼻咽癌的诊断筛查
Prokaryotic expression and preparation of GST/BFRF3 fusion protein of Epstein-Barr virus for screening nasopharyngeal carcinomas
摘要
Abstract
AIM:To construct a prokaryotic expression plasmid containing Epstein-Barr viral (EBV) capsidantigen BFRF3 gene and to observe the application of recombinant BFRF3 protein in the serological diagnosis of nasopharyngeal carcinoma (NPC).METHODS:DNA extracted from the B95-8 cells was used as the templates.Polymerase chain reaction (PCR) was used to generate a DNA fragment of BFRF3 gene,and a 531-bp DNA fragment was inserted into a PGEX-5X-1 vector.The recombinant plasmid was transformed into E.coli BL21 (DE3).The expression of GST/BFRF3 fusion protein was induced by IPTG,identified by both SDS-PAGE and Western blotting,and then purified by glutathionesepharose beads.The purified recombinant protein was coated to microplate for ELISA detection of EBV-IgA antibody in NPC patients.RESULTS:The GST/BFRF3 fusion protein was successfully expressed in E.coli.The molecular weight of the product was approximately 44 kD.The recombinant fusion protein GST/BFRF3 showed good immunoreactivity.A novel ELISA was established using GST/BFRF3 protein.Serum samples collected from the NPC patients and healthy controls were tested by this ELISA.The sensitivity and specificity of GST/BFRF3 tests for NPC patients were 65％ and 87％,respectively.CONCLUSION:The recombinant protein GST/BFRF3 is expressed in E.coli,and it has diagnostic value for screening of NPC patients.关键词
鼻咽肿瘤/EB病毒衣壳抗原/原核表达Key words
Nasopharyngeal neoplasms/Epstein-Barr viral capsid antigen/Prokaryotic expression分类
医药卫生引用本文复制引用
胡波,陈忠城,王东宁,李林..原核表达和制备EB病毒GST/BFRF3融合蛋白用于鼻咽癌的诊断筛查[J].中国病理生理杂志,2013,29(6):1147-1152,6.基金项目
教育部第44批留学回国人员科研启动基金[教外司留(2012)940] （2012）
