Selection and Identification of a DNA Aptamer Targeted to Vibrio parahemolyticus
详细信息 查看全文
- 作者：Nuo Duan ; Shijia Wu ; Xiujuan Chen ; Yukun Huang ; Zhouping Wang
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2012
- 出版时间：April 25, 2012
- 年：2012
- 卷：60
- 期：16
- 页码：4034-4038
- 全文大小：268K
- 年卷期：v.60,no.16(April 25, 2012)
- ISSN：1520-5118
文摘
A whole-bacterium systemic evolution of ligands by exponential enrichment (SELEX) method was applied to a combinatorial library of FAM-labeled single-stranded DNA molecules to identify DNA aptamers demonstrating specific binding to Vibrio parahemolyticus. FAM-labeled aptamer sequences with high binding affinity to V. parahemolyticus were identified by flow cytometric analysis. Aptamer A3P, which showed a particularly high binding affinity in preliminary studies, was chosen for further characterization. This aptamer displayed a dissociation constant (Kd) of 16.88 卤 1.92 nM. Binding assays to assess the specificity of aptamer A3P showed a high binding affinity (76%) for V. parahemolyticus and a low apparent binding affinity (4%) for other bacteria. Whole-bacterium SELEX is a promising technique for the design of aptamer-based molecular probes for microbial pathogens that does not require the labor-intensive steps of isolating and purifying complex markers or targets.