转基因玉米EPSPS蛋白适体的筛选与结构分析  

Screening and Structure Analysis of the Aptamers against the EPSPS Protein from Genetically Modified Maize (Zea mays)

作　　者：江树勋[1,4] 黄新[2] 邵碧英[1] 陈敏[1] 陈洪俊[1] Jun Sheng Lin 陈文炳[1] 吴亚君[2] 潘大仁[4] 

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,福州350001 [2]中国检验检疫科学研究院,北京100025 [3]Centre for Reproduction and Genomics, AgResearch, Invermay, New Zealand [4]福建农林大学生命科学学院,福州350002

出　　处：《热带作物学报》2010年第4期600-604,共5页Chinese Journal of Tropical Crops

基　　金：科技部转基因生物新品种培育科技重大专项(No.2009ZX08012-001B);中国检验检疫科学研究院基本科研业务费专项资金资助项目(No.2009JK001);福建省科技攻关项目(No.2008Y0001)资助

摘　　要：通过体外合成长度为70个碱基的随机ssDNA文库,采用指数富集配基的系统进化(SELEX)技术,以转基因玉米EPSPS蛋白为靶目标进行16轮筛选,将筛选到的适体群进行克隆、测序,初步获得与转基因玉米EPSPS蛋白特异性结合的适体。并用Macaw2.05和DNAMAN软件对每个适体的保守序列和二级结构进行分析。结果表明:一级结构分析序列同源性可分为四大群,其中A群有11条序列含有67个保守碱基;二级结构分析结果表明,茎环状和口袋状等结构可能是适体与转基因玉米EPSPS蛋白结合的结构基础。利用SELEX技术获得与转基因玉米EPSPS蛋白特异结合的适体群,其一级和二级结构与亲和力密切相关。Objective： To obtain aptamers that specially binds EPSPS protein from genetically modified maize （Zea mays）. Methods： A custom synthesized 113-mer random ssDNA library was subjected to 16 rounds of selection against recombinant EPSPS protein from genetically modified maize by using SELEX method. The selected aptamers were cloned and sequenced. Macaw 2.05 and DNAMAN package were employed to analyze the conserved sequences and secondary structure of the aptamers, respectively. Results： Primary structure analysis of sequence homology can be divided into four groups, of which there are 11 sequences containing 67 conserved bases in group A. The secondary structure analysis revealed possible stem-loops and pockets for binding to EPSPS protein. Conclusion： Aptamers against EPSPS protein had been obtained by SELEX method. The primary structure and the secondary structure are closely related to affinity.

关 键 词：转基因玉米 EPSPS蛋白 SELEX 适体 结构 

分 类 号：Q81[生物学—生物工程]