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| 靶标诱导变构解离SELEX技术筛选皮质醇特异核酸适体 | |
| 引用本文： | 张岭,王新兴,战锐,弓景波,钱令嘉. 靶标诱导变构解离SELEX技术筛选皮质醇特异核酸适体[J]. 军事医学科学院院刊, 2011, 35(10): 732-736 |
| 作者姓名： | 张岭 王新兴 战锐 弓景波 钱令嘉 |
| 作者单位： | 军医医学科学院基础医学研究所,全军军事认知与心理卫生研究中心,北京,100850 |
| 基金项目： | 国家科技重大专项课题(2008ZXJ090-011); 国家自然科学基金青年资助项目(81000584) |
| 摘 要： | 目的通过指数富集配体系统进化（SELEX）技术获得皮质醇特异性核酸适体,建立高品质、超敏、易用和稳定的皮质醇检测试剂盒。方法人工合成两端固定序列,中间为35 bp随机序列,生成80 bp长度ssDNA文库,捕获序列固定在磁珠上,文库与其退火组装,皮质醇的特异核酸适体通过我们建立的靶标诱导变构解离SELEX（TISSD-SELEX）技术获得。最后一轮的PCR产物经克隆测序,MegAlign和RNAstructure软件分析其一级和二级结构。实时（real-time）apta-PCR方法分析G12的亲和活性。结果经过12轮筛选后,获得10个皮质醇的核酸适体。序列分析显示,特异核酸适体以高G含量核酸序列为主,软件预测其中8个序列主要以G四联体二级结构为主,候选核酸适体G12显示了可以作为生物探针用于皮质醇检测。结论成功建立TISSD-SELEX技术,获得小分子靶标皮质醇核酸适体。小分子靶标无需将其偶联在固相介质或大分子上,对靶分子结构、性质无特殊要求,因而有望成为小分子靶标核酸适体筛选的通用技术方法。 |
| 关 键 词： | SELEX 皮质醇 核酸适体 纳米技术 |
Screening aptamers of cortisol by target-induced-structure-switching-dissociation SELEX |
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| ZHANG Ling,WANG Xin-xing,ZHAN Rui,GONG Jing-bo,QIAN Ling-jia. Screening aptamers of cortisol by target-induced-structure-switching-dissociation SELEX[J]. Bulletin of the Academy of Military Medical Sciences, 2011, 35(10): 732-736 | |
| Authors: | ZHANG Ling WANG Xin-xing ZHAN Rui GONG Jing-bo QIAN Ling-jia |
| Affiliation: | ZHANG Ling,WANG Xin-xing,ZHAN Rui,GONG Jing-bo*,QIAN Ling-jia* (Institute of Basic Medical Sciences,Research Center for Military Cognition and Mental Hygiene of PLA,Academy of Military Medical Sciences,Beijing 100850,China) |
| Abstract: | ObjectiveTo obtain and identify aptamers which can specifically bind to cortisol by systematic evolution of ligands by exponential enrichment(SELEX) protocol,in order to establish a high-quality,ultra-sensitive,simple & robust cortisol detection kit.MethodsA synthesized 80mer single stranded DNA random library containing 35 random sequences flanked by invariant primer was designed.Capture sequences were absorbed on the magnetic beads,the library and capture sequences were annealed and assembled,and the apta... |
| Keywords: | SELEX cortisol aptamer nanotechnology |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |