腐霉利核酸适配体的筛选与鉴定  

Screening and Identification of Procymidone Nucleic Acid Aptamers

作　　者：刘诺亚 高加乐 宋玉竹[1] 张金阳[1] 韩芹芹[1] LIU Nuoya;GAO Jiale;SONG Yuzhu;ZHANG Jinyang;HAN Qinqin(Faculty of Life Science and Technology,Kunming University of Science and Technology,Kunming 650500)

机构地区：[1]昆明理工大学生命科学与技术学院,云南昆明650000

出　　处：《分析科学学报》2023年第2期146-152,共7页Journal of Analytical Science

基　　金：国家自然科学基金(32160601);云南省基础研究计划-面上项目(202001AT070033)。

摘　　要：本文通过氧化石墨烯-体外指数富集配体系统进化技术,筛选并获得靶向腐霉利的核酸适配体Pr-A06和Pr-A08,通过二级结构预测,两种核酸适配体形成茎环结构和分子内G-四链体,结构稳定。使用胶体金法初步鉴定Pr-A06和Pr-A08,研究结果表明Pr-A06和Pr-A08解离常数(K d)分别为42.94±8.142 nmol/L、27.01±6.519 nmol/L,亲和力较好,并且特异性良好。Pr-A06浓度在0.1~1μg/mL之间有良好的线性范围,检测限为0.153μg/mL,Pr-A08浓度在0.05~1μg/mL之间有良好的线性范围,检测限为0.115μg/mL。本研究筛选的适配体为腐霉利的快速检测提供了新条件。In this work,nucleic acid aptamers Pr-A06 and Pr-A08 targeting putrescine were screened and obtained by a graphene oxide-systematic evolution of ligands by exponential enrichment(SELEX).The two nucleic acid aptamers formed stem-loop structures and intramolecular G-tetragonal with stable structures as predicted by secondary structures.The preliminary identification of Pr-A06 and Pr-A08 using the colloidal gold method showed good affinity and specificity with dissociation constant(K d)of 42.94±8.142 nmol/L and 27.01±6.519 nmol/L for Pr-A06 and Pr-A08,respectively.Pr-A06 showed good linearity between 0.1-1μg/mL with a limit of detection of 0.153μg/mL and Pr-A08 showed good linearity between 0.05-1μg/mL with a limit of detection of 0.115μg/mL.The aptamers screened in this study provide new tools for the rapid detection of putrescine.

关 键 词：核酸适配体 氧化石墨烯-SELEX技术 腐霉利 胶体金法 

分 类 号：Q503[生物学—生物化学]