食品科学 ›› 2011, Vol. 32 ›› Issue (13): 194-197.doi: 10.7506/spkx1002-6630-201113042

• 生物工程 • 上一篇    下一篇

应用SELEX技术筛选沙门氏菌抗原的适配子

郎春燕1,2,江树勋1,*,邵碧英1,陈文炳1,傅碧忠1,缪婷玉1,陈融斌3,黄晓蓉1   

  1. 1.福建出入境检验检疫局,福建省检验检疫技术研究重点实验室 2.福建农林大学食品科学学院 3.集美大学水产学院
  • 出版日期:2011-07-15 发布日期:2011-07-02
  • 基金资助:
    国家质检总局科技计划项目(2008IK164);福建省科技攻关计划重点项目(2008Y0001)

Screening of High-affinity DNA Aptamers to Salmonella Antigen by SELEX Technique

LANG Chun-yan1,2,JIANG Shu-xun1,*,SHAO Bi-ying1,CHEN Wen-bing1,FU Bi-zhong1,MIAO Ting-yu1, CHEN Rong-bin3,HUANG Xiao-rong1   

  1. 1. Fujian Key Laboratory for Technology Research of Inspection and Quarantine, Fujian Entry-Exit Inspection and Quarantine Bureau, Fuzhou 350001, China;2. College of Food Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 3. Fisheries College of Jimei University, Xiamen 361021, China
  • Online:2011-07-15 Published:2011-07-02

摘要: 目的:应用指数富集配体系统进化(SELEX)技术筛选沙门氏菌抗原的高亲和性适配子。方法:首先合成一个全长78个核苷酸中间含35个随机序列的随机单链寡核苷酸序列(ssDNA)文库,再以环氧乙烷丙烯酸珠子作为筛选介质,利用生物素-抗地高辛碱性磷酸酶显色系统检测DNA适配子与沙门氏菌抗原的亲和力,以获得沙门氏菌抗原的高亲和性适配子。结果:随着筛选轮数的增加,DNA适配子与沙门氏菌抗原结合后显色,吸光度逐渐增加,初步获得了沙门氏菌抗原的高亲和性适配子。结论:实验所用的筛选流程是适当的,可以考虑推广用于类似靶目标的筛选。

关键词: 指数富集配体系统进化技术, 沙门氏菌, 抗原, 适配子

Abstract: Objective: To screen high-affinity aptamers to Salmonella antigen by applying SELEX (system evolution of ligands by exponential enrichment) method and establish a rapid detection method for Salmonella in food. Methods: First, a single stranded DNA (ssDNA) random library with 78 nucleotides in length containing 35 random sequences was constructed. Subsequently, Eupergit C was used as the screening medium in the SELEX method. The binding of aptamers to Salmonella antigen was visualized by Biotin-Streptavidin-Anti-Digoxigenin-AP system to screen high-affinity aptamers out. Results: The binding assay demonstrated a gradual decrease in the absorbance at 405 nm as the number of screening rounds increased. High-affinity aptamers binding to Salmonella antigen were successfully obtained from the initial random ssDNA library. Conclusion: The screening process is simple and can be used for similar targets.

Key words: system evolution of ligands by exponential enrichment (SELEX), Salmonella, antigen, aptamer

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