高等学校化学学报

高等学校化学学报

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采用毛细管电泳技术筛选特异识别蓖麻毒素适配子的研究

唐吉军1, 谢剑炜1, 邵宁生2, 郭磊1, 闫妍1   

    1. 军事医学科学院毒物药物研究所,
    2. 基础医学研究所, 北京 100850
  • 收稿日期:2005-10-28 修回日期:1900-01-01 出版日期:2006-10-10 发布日期:2006-10-10
  • 通讯作者: 谢剑炜

Capillary Electrophoresis as a Tool for Screening Aptamer with High Affinity and High Specificity to Ricin

TANG Ji-Jun1, XIE Jian-Wei1, SHAO Ning-Sheng2, GUO Lei1, YAN Yan1   

    1. Beijing Institute of Pharmacology & Toxicology,
    2. Beijing Institute of Basic Medical Sciences, Beijing 100850, China
  • Received:2005-10-28 Revised:1900-01-01 Online:2006-10-10 Published:2006-10-10
  • Contact: XIE Jian-Wei

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被引次数

22

摘要： 采用指数富集配基的系统进化(SELEX)技术从随机寡核苷酸文库中筛选获得特异识别蓖麻毒素靶分子的适配子. 将毛细管电泳技术作为分离手段引入到SELEX筛选中, 利用毛细管电泳高效的分离能力使得筛选周期大大缩短. 酶联免疫和斑点杂交实验结果表明, 仅经4轮筛选即可获得特异识别蓖麻毒素蛋白的寡核苷酸适配子.

关键词: 指数富集配基的系统进化, 毛细管电泳, 随机单链DNA文库, 蓖麻毒素

Abstract: SELEX(Systematic Evolution of Ligands by Exponential Enrichment) process allows a rapid isolation of oligonucleotide sequences, known as aptamers binding target molecules with high affinity and specificity, which are equal or superior to those of antibodies, from a very large nucleic acid library. In this paper, capillary electrophoresis(CE) as a separation approach was introduced to SELEX procedure. The great high efficiency of CE gives rise to greatly shorten the selection procedure. The results from enzyme-linked assay and dot blot experiment show that an enrichment pool was obtained after four rounds selection, which can specifically be used to recognize ricin.

Key words: Systematic evolution of ligands by exponential enrichment(SELEX), Capillary electrophoresis(CE), Single-stranded DNA(ssDNA) pool, Ricin

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