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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。
A randomized ssDNA library with a length of 70 bases was synthesized in vitro. Sixteen rounds of screening were carried out using the SELEX (exponential enrichment ligand) technique and EPSPS target of transgenic corn as the target. The selected aptamers were cloned , Sequenced and initially obtained aptamers that specifically bind to the EPSPS protein of the transgenic maize. The conserved sequence and secondary structure of each aptamer were analyzed using Macaw 2.05 and DNAMAN software. The results showed that the primary structure analysis sequence homology can be divided into four groups, of which 11 sequences in group A contained 67 conserved bases. The secondary structure analysis showed that the ring-shaped and pocket-like structures of stems may be aptamers Structural basis for binding to EPSPS protein from transgenic corn. The use of SELEX technology to obtain EPSPS protein and transgenic corn specific binding aptamer group, the primary and secondary structure and affinity are closely related.