运用SELEX技术筛选嗜水气单胞菌适配子

Screening of the DNA Aptamers Against Aeromonas hydrophila by Using SELEX Method

以嗜水气单胞菌为研究对象,采用适体筛选技术(SELEX,system evolution of ligands by expo-nential enrichment),利用体外合成的长度为78个核苷酸的随机ssDNA文库,以嗜水气单胞菌为靶目标进行12轮的筛选,利用生物素-抗地高辛碱性磷酸酶显色系统检测DNA适配子与嗜水气单胞菌的亲和力.研究结果表明:随着筛选轮数的增加,DNA适配子与嗜水气单胞菌结合后显色,A(absorbance)值逐渐增加,亲和力由最初的0.32升到第9轮的0.72.研究已初步筛选到与嗜水气单胞菌具有高亲和力的DNA适配子,为嗜水气单胞菌的二级结构研究及其检测试剂盒的研发提供了基础.

To obtain the aptamers of Aeromonas hydrophila, a custom synthesized 78 -mer random ssDNA library was subjected to 12 rounds of selection against Aeromonas hydrophila by using SELEX method. The selected aptamers were cloned and sequenced. Macaw 2. 05 and DNAMAN package were employed to analyze the conserved sequences and secondary structure of the aptamers, respectively. The results showed that the binding assay demonstrated that the absorbance at 405 nm generally increased from 0. 32 to 0. 92. The results indicated that aptamers with high affinity against Aeromonas hydrophila had been obtained by SEL- EX methods and provided a basis for its secondary structure and development of detection kit.