原核表达HPV16 L1蛋白ELISA方法检测血清抗体  

ELISA detection of prokaryotic expression serum antibody of HPV16 L1 protein

作　　者：赵丽晶[1] 王学林[2] 许多[1] 梁作文[1] 郭恒[2] 孙树民[2] 赵淑华[3] 赵雪俭[1] 郑晶莹[3] 

机构地区：[1]吉林大学基础医学院病理生理学教研室,吉林长春130021 [2]吉林大学人兽共患病教育部重点实验室 [3]吉林大学第二医院妇产科

出　　处：《中国妇幼保健》2010年第3期395-397,共3页Maternal and Child Health Care of China

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。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.

关 键 词：HPV16 L1蛋白 原核表达 ELISA检测 

分 类 号：R737.3[医药卫生—肿瘤]