阪崎肠杆菌特异性DNA适配子的SELEX筛选研究.pdfVIP

中国科技论文在线 阪崎肠杆菌特异性 DNA 适配子的 SELEX 筛选研究# 韩晓晓，夏雨，王周平** 5 （江南大学食品学院，江苏无锡 214122 ） 摘要：阪崎肠杆菌是一种条件性致病菌，对于婴幼儿的影响重大。本研究应用体外指数富集 配体的系统进化（SELEX）技术和流式细胞分析筛选获得 4 条与阪崎肠杆菌具有高亲和力的 荧光DNA适配子。其中，B25P的解离常数为 38.61±5.624 nM，对阪崎肠杆菌显示了高亲和 性，对其他细菌的亲和力低（不超过15%），具有良好的特异性，是进一步应用的候选序列。 10 全菌SELEX技术无需分离纯化目标的繁琐操作，为基于适配子探针的致病菌检测提供了现实 基础。 关键词：适配子；阪崎肠杆菌；SELEX技术；流式细胞术 中图分类号：TS201.6；TS201.2 15 Selection and Identification of a DNA Aptamer Targeted to Cronobacter Sakazakii HAN Xiaoxiao, XIA Yu, WANG Zhouping (School of Food Science and Technolohy, Jiangnan University, Jiangsu WuXi 214122) Abstract: Cronobacter sakazakii(C.sakazakii ) is a kind of conditional pathogenic bacteria which 20 has a great influence for infants. Four FAM-labeled DNA aptamers demonstrating high binding affinity to Cronobacter sakazakii were identified by a whole-bacterium systemic evolution of ligands by exponential enrichment (SELEX) method and flow cytometric analysis. Aptamer B25P was chosen for further characterization. It displayed a dissociation constant of 38.61±5.624 nM and a high binding affinity .The fluorescence intensity was 81.86% for C. sakazakii and less than 25 15% for other bacteria, which also characterize its high specificity. Whole-bacterium SELEX is a promising technique for the detection of microbial pathogens that does not require the labor-intensive steps of isolating and purifying complex markers or targets. Keywords: Aptamer; Cronobacter sakazakii; SELEX; Flow Cytometr