应用SELEX技术筛选沙门氏菌抗原的适配子  

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

作　　者：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] 

机构地区：[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,福建福州350001 [2]福建农林大学食品科学学院,福建福州350002 [3]集美大学水产学院,福建厦门361021

出　　处：《食品科学》2011年第13期194-197,共4页Food Science

基　　金：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)

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。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.

关 键 词：指数富集配体系统进化技术 沙门氏菌 抗原 适配子 

分 类 号：Q789[生物学—分子生物学]