单核增生李斯特氏菌适体的筛选与结构分析  

Screening and Structural Analysis of Aptamer Binding to Listeria monocytogenes

作　　者：陈敏[1,2] 江树勋[1] 邵碧英[1] 高如承[2] 陈文炳[1] 陈彬[1] 李寿崧[1] 缪婷玉[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]福建师范大学生命科学学院,福建福州350108

出　　处：《食品科学》2009年第19期197-199,共3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);福建检验检疫局科技计划项目(FK2006-13)

摘　　要：为获得与单核增生李斯特氏菌特异性结合的适体,对体外合成长度为78个碱基的随机ssDNA文库,采用SELEX技术,以单核增生李斯特氏菌为靶标进行10轮筛选。将筛选到的适体群进行克隆、测序,用Macaw2.05和DNAMAN软件对每一适体的保守序列和二级结构进行分析。一级结构分析获得6个保守序列:TTTTTTT、TGCTCTT、CGGGGGT、GTTTTT、XTGTT、XTTGT,其中2号和12号、7号和9号、158、159和160号适体序列完全一致。二级结构分析表明,茎环状和口袋状等结构可能是适体与单核增生李斯特氏菌结合的结构基础,其一级和二级结构与亲和力密切相关。Objective： To obtain aptamer that specially binds Listeria monocytogenes. Methods： A custom synthesized 78-mer random ssDNA library was subjected to 10 rounds of selection against Listeria monocytogenes by SELEX method. The selected aptamers were cloned and sequenced. Macaw 2.05 and DNAMAN package were employed to analyze conserved sequences and secondary structure of aptamers, respectively. Results： 6 conserved sequences such as TITTTTT, TGCTCTT, CGGGGGT, GTITTT, XTGTT and XTTGT were obtained. Absolutely identical sequences were exhibited in the pairs of NO. 2 and 12, 7 and 9, 158, 159 and 160 aptamer. Secondary structure analysis revealed possible stem-loops and pockets for binding to Listeria monocytogenes. Conclusion： Aptamers with binding capability to Listeria monocytogenes have been obtained by SELEX methods. Their affinities are closely related to the primary structure and the secondary structure.

关 键 词：SELEX 单核增生李斯特氏菌 适体 一级结构 二级结构 

分 类 号：Q789[生物学—分子生物学]