CAJ | 학술논문

目的筛选识别甲胺磷的 DNA 适体.方法体外合成 ssDNA 文库,将其固相化于琼脂糖凝胶颗粒表面,液相中甲胺磷与其结合的 DNA 分子从固相表面分离,进而建立非固相化 SELEX 技术,最终获得识别甲胺磷的适体,采用 DNAMAN 和 RNA structure 软件对适体进行一、二级结构分析.结果经过10轮筛选, ssDNA 文库与甲胺磷的亲和力呈上升趋势,随机挑选的20个阳性克隆适体根据一级结构的同源性可分为5个家族,二级结构预测以茎环结构为主.结论本方法大大提高了适体的筛选效率,最终筛选得到能识别甲胺磷的 DNA 适体.
목적사선식별갑알린적 DNA 괄체.방법체외합성 ssDNA 문고,장기고상화우경지당응효과립표면,액상중갑알린여기결합적 DNA 분자종고상표면분리,진이건립비고상화 SELEX 기술,최종획득식별갑알린적괄체,채용 DNAMAN 화 RNA structure 연건대괄체진행일、이급결구분석.결과경과10륜사선, ssDNA 문고여갑알린적친화력정상승추세,수궤도선적20개양성극륭괄체근거일급결구적동원성가분위5개가족,이급결구예측이경배결구위주.결론본방법대대제고료괄체적사선효솔,최종사선득도능식별갑알린적 DNA 괄체.
@@@@Objective To select aptamers against methamidophos. Methods The ssDNA pool which was synthesized in vitro was immoboilized on surface of agarose gel. The ssDNAs that bound methamidophos were released from surface of agarose gel. Methamidophos aptamers were selected by non-solid phase SELEX. The primary and secondary structure of methamidophos aptamers were analysed by DNAMAN and RNA structure software. Results Affinity between ssDNA pool and methamidophos was increased after 10 selection rounds. The 20 ssDNAs which were positive clones were divided into five groups based on primary sequence homology. Hairpin loop was the main motif in the predicted secondary structure and it was supposed to be the binding part of the aptamers to methamidophos. Conclusion The efficiency of screening of the aptamer was greatly im-proved using this method and the methamidophos-specific DNA aptamers were selected successfully.