首页 | 期刊简介 | 编委会 | 投稿指南 | 常用下载 | 联系我们 | 期刊订阅 | In English
引用本文:林筱钧,许净,郑江,鄢庆枇,高得骥,范云庭,黄将远.变形假单胞菌(Pseudomonas plecoglossicida)核酸适配体的SELEX筛选.海洋与湖沼,2023,54(2):583-590.
【打印本页】   【HTML】   【下载PDF全文】   查看/发表评论  下载PDF阅读器  关闭
←前一篇|后一篇→ 过刊浏览    高级检索
本文已被:浏览 428次   下载 620 本文二维码信息
码上扫一扫!
分享到: 微信 更多
变形假单胞菌(Pseudomonas plecoglossicida)核酸适配体的SELEX筛选
林筱钧1,2, 许净1, 郑江1,2, 鄢庆枇1, 高得骥1, 范云庭1, 黄将远1
1.集美大学水产学院 福建厦门 361021;2.鳗鲡现代产业技术教育部工程研究中心 福建厦门 361021
摘要:
变形假单胞菌(Pseudomonas plecoglossicida)是大黄鱼内脏白点病的主要致病菌。核酸适配体,因其亲和力高、特异性好等多个优点,在微生物检测等多个领域都有较好的应用前景。以变形假单胞菌为靶目标,采用SELEX筛选方法,从高频序列中筛选获得了变形假单胞菌的5个核酸适配体,测定了这些核酸适配体对变形假单胞菌的亲和特异性及其亲和常数(Kd)和最大亲和力(Am),并对其二级结构进行了分析研究。结果表明,这5个核酸适配体对变形假单胞菌的亲和力是其他非目标菌(嗜水气单胞菌、溶藻弧菌、哈维氏弧菌、大肠杆菌、迟钝爱德华氏菌、铜绿假单胞菌)的10倍以上,体现出了较好的亲和特异性。测定了5个核酸适配体(#1、#2、#3、#4、#5)的KdAm,相应的Kd值分别为(25.77±6.11)、(7.14±1.86)、(19.59±3.01)、(83.22±22.45)、(34.31±8.76) nmol/L,Am值分别为(789.23±38.46)、(1392.27±58.03)、(3012.50±106.19)、(1457.99±171.32)、(1070.91±70.16) nmol/L。二级结构模拟发现核酸适配体对靶标的结合能力可能主要与其二级结构中的大中型环有关。相关研究对于变形假单胞菌的快速检测及其病害防控都具有重要意义。
关键词:  变形假单胞菌  核酸适配体  亲和常数  最大亲和力  二级结构
DOI:10.11693/hyhz20220700194
分类号:Q939;S917.1
基金项目:福建省自然科学基金项目,2021J01823号;鳗鲡现代产业技术教育部工程研究中心开放基金项目,RE202205号,RE202308号;福建省特种水产配合饲料重点实验室开放课题项目,TMKJZ1909号。
附件
SELECTION OF APTAMERS OF PSEUDOMONAS PLECOGLOSSICIDA BY SELEX
LIN Xiao-Jun1,2, XU Jing1, ZHENG Jiang1,2, YAN Qing-Pi1, GAO De-Ji1, FAN Yun-Ting1, HUANG Jiang-Yuan1
1.Fisheries College of Jimei University, Xiamen 361021, China;2.Engineering Research Center of the Modern Technology for Eel Industry, Ministry of Education, Xiamen 361021, China
Abstract:
Pseudomonas plecoglossicida is the main pathogen of visceral white spot disease in large yellow croaker. Aptamer shows good application prospects in microbial detection due to many advantages such as high affinity and good specificity. Five aptamers of P. plecoglossicida were selected from the high-frequency sequences by SELEX. The affinities of the five aptamers towards P. plecoglossicida were more than 10 times those of the non-target bacteria (Aeromonas hydrophila, Vibrio alginolyticus, Vibrio harveyi, Escherichia coli, Edwardsiella tarda and Pseudomonas aeruginosa), which proved that the aptamers had good affinities and specificities. The affinity constants (Kd) and maximum affinities (Am) of the five aptamers (#1, #2, #3, #4, #5) were determined, and their Kd were (25.77±6.11), (7.14±1.86), (19.59±3.01), (83.22±22.45), (34.31±8.76) nmol/L, respectively, and their Am were (789.23±38.46), (1 392.27±58.03), (3 012.50±106.19), (1 457.99±171.32), and (1 070.91±70.16) nmol/L, respectively. Finally, the secondary structures of the aptamers were analyzed. It was found that the binding ability of aptamer with its target was related mainly to the large and medium rings in the secondary structure. The relative researches shall have important significance for the rapid detection of P. plecoglossicida and the control of disease caused by the pathogen.
Key words:  Pseudomonas plecoglossicida  aptamer  affinity constant  maximum affinity  secondary structure
版权所有 海洋与湖沼 Oceanologia et Limnlolgia Sinica Copyright©2008 All Rights Reserved
主管单位:中国科协技术协会 主办单位:中国海洋湖沼学会
地址:青岛市海军路88号  邮编:266400  电话:0532-82898753  E-mail:ols@qdio.ac.cn
技术支持:北京勤云科技发展有限公司