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余庆,刘明珠,李梦梦,黄帅帅,徐凤巧,李鹏飞.特异性检测卵形鲳鲹源哈维氏弧菌的核酸适配体的筛选与鉴定[J].广西科学院学报,2021,37(2):75-84. [点击复制]
- YU Qing,LIU Mingzhu,LI Mengmeng,HUANG Shuaishuai,XU Fengqiao,LI Pengfei.Selection and Identification of Aptamers Specifically Recognizing Pathogenic Vibrio harveyi Isolated from Cultured Golden Pompano (Trachinotus ovatus)[J].Journal of Guangxi Academy of Sciences,2021,37(2):75-84. [点击复制]
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| 特异性检测卵形鲳鲹源哈维氏弧菌的核酸适配体的筛选与鉴定 |
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余庆1, 刘明珠1, 李梦梦2, 黄帅帅1,3, 徐凤巧1, 李鹏飞1,3
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| (1.广西科学院, 广西壮族自治区渔业重大疫病防控与高效健康养殖产业技术工程研究中心, 广西海洋天然产物与组合生物合成化学重点实验室, 广西南宁 530007;2.河南师范大学生命科学学院, 河南新乡 453007;3.北部湾大学海洋学院, 广西北部湾海洋生物多样性养护重点实验室, 广西钦州 535011) |
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| 摘要: |
| 哈维氏弧菌（Vibrio harveyi）是海水养殖动物常见的主要致病菌之一，严重危害着我国华南沿海地区水产养殖业的健康可持续发展。针对哈维氏弧菌的快速检测诊断技术和抑菌技术等方面进行系统研究，并开发具有市场发展潜力的快速检测试剂盒和抗菌功能产品，对控制哈维氏弧菌的危害极其重要。为此，本研究以哈维氏弧菌为靶标，利用指数富集的配基系统进化（Systematic Evolution of Ligands by Exponential Enrichment，SELEX）技术，筛选特异性识别哈维氏弧菌的核酸适配体，并对核酸适配体的性质进行系统性研究。结果显示，核酸适配体H10、H13能高特异性和高亲和性地识别哈维氏弧菌，并且无毒副作用。同时，靶标性质分析表明核酸适配体（H10，H13）的靶标可能是细胞膜蛋白或膜蛋白相关成分。本研究运用SELEX技术筛选获得的ssDNA核酸适配体H10和H13，具有高特异性和亲和性，可用于研发操作便捷、灵敏度高的哈维氏弧菌快速检测技术。 |
| 关键词: 水产养殖 哈维氏弧菌 核酸适配体 特异性强 快速检测技术 |
| DOI：10.13657/j.cnki.gxkxyxb.20210816.001 |
| 投稿时间：2021-05-07 |
| 基金项目:国家自然科学基金项目（U20A20102），广西重点研发计划项目（桂科AB18221111），广西自然科学基金项目（2020GXNSFBA297161，2019GXNSFAA245054和AD19245022）和广西科学院基本科研业务费项目（2018YJJ903，2019YJJ1005，2018YJJ902）资助。 |
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| Selection and Identification of Aptamers Specifically Recognizing Pathogenic Vibrio harveyi Isolated from Cultured Golden Pompano (Trachinotus ovatus) |
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YU Qing1, LIU Mingzhu1, LI Mengmeng2, HUANG Shuaishuai1,3, XU Fengqiao1, LI Pengfei1,3
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| (1.Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Engineering Research Center for Fishery Major Diseases Control and Efficient Healthy Breeding Industrial Technology, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China;2.College of Life Science, Henan Normal University, Xinxiang, Henan, 453007, China;3.Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation, College of Marine Sciences, Beibu Gulf University, Qinzhou, Guangxi, 535011, China) |
| Abstract: |
| Vibrio harveyi is one of the main pathogenic bacteria that are common in marine aquaculture animals, which seriously endangers the healthy and sustainable development of the aquaculture industry in the coastal areas of South China. Systematic research on the rapid detection and diagnosis technology and antibacterial technology against V. harveyi, and the development of rapid detection kits and antibacterial products with market development potential are extremely important to control the harm of V. harveyi. This study took V. harveyi as the target and used the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology to screen for aptamers that specifically recognize V. harveyi, and systematically study the properties of aptamers. The results showed that the selected aptamers H10 and H13 could recognize target V. harveyi with high specificity and affinity without cytotoxicity. The target properties analysis results showed that the targets of aptamers H10 and H13 might be cell membrane proteins or membrane proteins related components. The ssDNA aptamers H10 and H13 selected by SELEX technology have high specificity and affinity targeting V. harveyi, which could be used to develop the rapid detection technology against V. harveyi with convenient operation and high sensitivity. |
| Key words: aquaculture Vibrio harveyi aptamer high specificity rapid detection assay |
