【摘要】 目的:从感染人乳头瘤病毒(HPV)的新鲜宫颈癌组织中扩增HPV16型晚期蛋白L1基因全长,构建原核表达载体,表达并纯化蛋白用于ELISA检测。方法:根据基因序列设计一对特异引物,用PCR的方法从宫颈癌组织DNA中获得L1基因,以pet28a为载体构建表达质粒,IPTG诱导表达蛋白,DEAE及葡聚糖凝胶分离纯化目的蛋白,将蛋白包被平底96孔板用于ELISA检测。结果:从宫颈癌临床标本克隆到HPV16型L1蛋白编码序列,其原核表达产物纯化后可用于ELISA检测。结论:成功获得人有抗原性的HPV16L1蛋白,可用于ELISA检测。更多还原

【Abstract】 Objective:To amplify the HPV16 late protein L1 gene in fresh cervical cancer tissues infected by human papillomavirus (HPV),construct prokaryotic expression vector,then the protein was expressed,purified and used for ELISA detection.Methods:Specific primers were designed according to gene sequence.L1 gene was obtained from cervical cancer tissues DNA by PCR to construct recombinant prokaryotic expression vector pet28a-HPV16-L1.L1 protein expression was induced by IPTG,purified by DEAE and sephadex,and packaged to flat-bottomed 96-well plates for ELISA detection.Results:HPV16 L1 protein coding sequence was amplified from cervical cancer samples,the prokaryotic expressing protein was purified and used for ELISA detection.Conclusion:HPV16 L1 protein is obtained successfully,which can be used for ELISA detection.更多还原