Isolation and characterization of DNA aptamers against Escherichia coli using a bacterial cell-systematic evolution of ligands by exponential enrichment approach
Isolation and characterization of DNA aptamers against Escherichia coli using a bacterial cell-systematic evolution of ligands by exponential enrichment approach
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DOI：
10.1016/j.ab.2013.01.014
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发表时间：
2013-05-01
影响因子：
2.9
通讯作者：
Kim, Byoung Chan
中科院分区：
文献类型：
--
作者：
Kim, Yeon Seok;Song, Min Young;Kim, Byoung Chan
适体是针对各种靶标（如蛋白质、小有机化合物、金属离子甚至细胞）的强大捕获探针。在这项研究中，我们分离和特点的单链DNA（ssDNA）的适体对大肠杆菌。在使用细菌细胞-SELEX（通过指数富集的配体系统进化）方法进行10轮选择后，总共分离出28个ssDNA。其他细菌物种（肺炎克雷伯氏菌、弗氏柠檬酸杆菌、产气肠杆菌和表皮葡萄球菌）用于反选择以增强ssDNA适体对E.杆菌最后，4个ssDNA适体对大肠杆菌表现出较高的亲和性和选择性。coli的解离常数（K-d）。大肠杆菌的浓度范围估计为12.4至25.2 nM。这些适体不与其他细菌物种结合，包括四种计数细胞，但它们对不同的E.大肠杆菌菌株。这四种适体与E.用荧光显微镜直接观察大肠杆菌。(C)2013 Elsevier Inc. All rights reserved.
Aptamers are powerful capturing probes against various targets such as proteins, small organic compounds, metal ions, and even cells. In this study, we isolated and characterized single-stranded DNA (ssDNA) aptamers against Escherichia coli. A total of 28 ssDNAs were isolated after 10 rounds of selection using a bacterial cell-SELEX (systematic evolution of ligands by exponential enrichment) process. Other bacterial species (Klebsiella pneumoniae, Citrobacter freundii, Enterobacter aerogenes, and Staphylococcus epidermidis) were used for counter selection to enhance the selectivity of ssDNA aptamers against E. coli. Finally, four ssDNA aptamers showed high affinity and selectivity to E. coli, The dissociation constants (K-d) of these four ssDNA aptamers to E. coli were estimated to range from 12.4 to 25.2 nM. These aptamers did not bind to other bacterial species, including four counter cells, but they showed affinity to different E. coli strains. The binding of these four aptamers to E. coli was observed directly by fluorescence microscopy. (C) 2013 Elsevier Inc. All rights reserved.