| 溶藻弧菌适配子亲和力测定条件的优化及其特异性研究 |
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| 引用本文： | 都聚敏,李晶蕊,汤学敏,郑江,黎中宝,鄢庆枇.溶藻弧菌适配子亲和力测定条件的优化及其特异性研究[J].安徽农业科学,2011,39(32):19874-19876. |
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| 作者姓名： | 都聚敏 李晶蕊 汤学敏 郑江 黎中宝 鄢庆枇 |
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| 作者单位： | 集美大学水产学院,福建省高校水产科学技术与食品安全重点实验室,福建厦门361021 |
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| 基金项目： | 福建省教育厅科技项目(JB10096);集美大学科研启动基金(ZQ2011002);集美大学创新团队项目(2010A004);集美大学2011年度学生科研基金项目 |
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| 摘 要： | 目的]优化溶藻弧菌适配子亲和力的测定条件,并验证适配子的亲和特异性。方法]采用地高辛-过氧化物酶显色系统,通过正交试验确定亲和力测定的适宜条件,并在该条件下进行特异性验证。结果]优化后亲和力的测定条件为：菌浓度3.0×10^8个/ml,结合时间40 min,结合温度28℃;经验证,该适配子对溶藻弧菌具有较高的亲和特异性。结论]获得了溶藻弧菌适配子亲和力测定的优化条件,并证明了该适配子与溶藻弧菌具有较好的亲和特异性,为溶藻弧菌的SELEX筛选及其快速检测提供了参考。
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| 关 键 词： | SELEX 溶藻弧菌 适配子 亲和力 特异性 |
Optimization of Affinity Determination Condition of Aptamer for Vibrto alginolyticus and Specificity Analysis |
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| Institution: | HAO Ju-min et al(Key Laboratory of Science and Technology for Aquaculture and Food Safety,Fisheries College of Jimei University,Xiamen,Fujian 361021) |
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| Abstract: | Objective] The aim was to optimize the condition for the affinity detection of an aptamer against Vibrio alginolyticus and to validate the aptamer′s specific affinity to the microorganism.Method] Based on the digoxigenin-peroxidase color system,the optimal condition was obtained by the orthogonal experiment and the specific affinity of the aptamer was validated under the optimized conditions.Result] The optimum condition for the affinity detection of the aptamer was obtained as follows:bacteria concentration 3.0×108 cell/ml,binding time 40 min,and binding temperature 28 ℃;the aptamer was validated to have the specific affinity to the microorganism Vibrio alginolyticus under the optimum condition.Conclusion] The optimum condition for the affinity detection of the aptamer against Vibrio alginolyticus was obtained,and the aptamer was proved to have good specific affinity to the microorganism,which would be a valuable reference for the SELEX of aptamers against Vibrio alginolyticus and the quick detection of the microorganism. |
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| Keywords: | SELEX Vibrio alginolyticus Aptamer Affinity Specificity |
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