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| 原核表达HPV16 L1蛋白ELISA方法检测血清抗体 | |
| 引用本文： | 赵丽晶,王学林,许多,梁作文,滕菲,郭恒,孙树民,赵淑华,赵雪俭. 原核表达HPV16 L1蛋白ELISA方法检测血清抗体[J]. 中国妇幼保健, 2010, 25(35) |
| 作者姓名： | 赵丽晶 王学林 许多 梁作文 滕菲 郭恒 孙树民 赵淑华 赵雪俭 |
| 作者单位： | 1. 吉林大学白求恩医学院病理生理学系,吉林,长春,130021 2. 吉林大学人兽共患病教育部重点实验室 3. 吉林大学白求恩第二医院妇产科 |
| 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HPV16 L1蛋白,可用于ELISA检测。 |
| 关 键 词： | 人乳头瘤病毒 原核表达 血清抗体 |
Detection of serum antibody of prokaryotic expression HPV16 L1 protein by ELISA |
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| Abstract: | Objective:To amplify the full-length of HPV16 late protein L1 in fresh cervical cancer tissues infected by human papillomavirus (HPV), construct prokaryotic expression vector, express and purify the protein for ELISA detection.Methods:A pair of specific primer was designed according to gene sequence.PCR method was used to obtain L1 gene from cervical cancer tissues DNA, pet28a was used as vector to construct prokaryotic expression plasmid, L1 protein was induced by IPTG, separated and purified by DEAE and sephadex, then L1 protein was packaged to flat-bottomed 96-well plates for ELISA detection.Results:The prokaryotic expression products were used for ELISA detection after separation and purification by cloning clinical samples of cervical cancer to HPV16 L1 protein coding sequence.Conclusion:HPV16 L1 protein is obtained successfully, which can be used for ELISA detection. |
| Keywords: | Human papillomavirus Prokaryotic expression Serum antibody |
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