Selection and characterization of single stranded DNA aptamers recognizing fumonisin B1
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- 作者：Xiujuan Chen (1)
Yukun Huang (1)
Nuo Duan (1)
Shijia Wu (1)
Yu Xia (1)
Xiaoyuan Ma (1)
Changqing Zhu (2)
Yuan Jiang (2)
Zhansheng Ding (1)
Zhouping Wang (1) - 关键词：Fumonisin B1 ; Aptamer ; SELEX ; Magnetic beads ; Competition
- 刊名：Microchimica Acta
- 出版年：2014
- 出版时间：August 2014
- 年：2014
- 卷：181
- 期：11-12
- 页码：1317-1324
- 全文大小：720 KB
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Yukun Huang (1)
Nuo Duan (1)
Shijia Wu (1)
Yu Xia (1)
Xiaoyuan Ma (1)
Changqing Zhu (2)
Yuan Jiang (2)
Zhansheng Ding (1)
Zhouping Wang (1)
1. State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China
2. Animal, Plant and Food Inspection Centre, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001, China - ISSN：1436-5073
文摘
We present an improved method for the selection of single-stranded DNA aptamers that can recognize fumonisin B1 (FB1). FB1 is a carcinogenic mycotoxin mainly found in corn and corn-based food products worldwide, posing a global threat to feed and food safety. Selection was based on the mag-SELEX (magnetic bead systematic evolution of ligands by exponential enrichment) technology modified by adopting free analogs of targets rather than immobilized targets for counter selections. Firstly, aptamer candidates for FB1 were selected from an 80?nt random DNA library after 13 rounds of selection. Next, binding assays were performed for affinity evaluation, and circular dichroism spectroscopy was used to investigate their conformation. A high-affinity aptamer designated as F10 (with a dissociation constant of 62?±- nM) was identified and tested for its specificity by competitive binding assays. The results demonstrate that this improved mag-SELEX technology facilitates aptamer screening because it avoids the tedious immobilization of counter-selection molecules on magnetic beads. The aptamers obtained by this technique open new possibilities for the detection of FB1 via aptasensors. Graphical Abstract Illustration of SELEX selection procedure. An improved method for selection of ssDNA aptamers recognizing fumonisin B1 was developed based on magnetic beads. Free analogs of targets rather than immobilized were used for counter selections, facilitating the selection and leading to affinity-raised aptamers.