芝麻过敏原Ses i 2核酸适体的筛选与鉴定研究  

Screening and Identification of Nucleic Acid Aptamer for the Sesame Allergen Ses i 2

作　　者：李洋[1,2] 于宁[2] 康文瀚[2] 张九凯 杜欣军[1] 陈颖[2] LI Yang;YU Ning;KANG Wenhan;ZHANG Jiukai;DU Xinjun;CHEN Ying(College of Food Science and Engineering,Tianjin University of Science and Technology,Tianjin 300457;Chinese Academy of Inspection and Quarantine,Beijing 100176)

机构地区：[1]天津科技大学食品科学与工程学院,天津300457 [2]中国检验检疫科学研究院,北京100176

出　　处：《核农学报》2025年第1期59-67,共9页Journal of Nuclear Agricultural Sciences

基　　金：中国检验检疫科学研究院基本科研业务费项目(2022JK04)。

摘　　要：芝麻是八大类食物过敏原之一,快速准确识别芝麻过敏原对预防其过敏有重要意义。核酸适配体可以高效识别靶标过敏原,在过敏原检测中有良好的应用前景。为了获得芝麻主要过敏原Ses i 2的特异性核酸适体,本研究以Ses i 2为靶标,通过磁珠筛选法(磁珠-SELEX)开展10轮筛选,经由高通量测序获得6条候补序列(S1~S6),并进行家族性、同源性分析及二级结构预测。结果表明,6条候选核酸适体的重复率可达46.38%,其自由能在-9.02到-2.47 kcal·moL^(-1)之间,根据自由能能量稳定原则,S1和S5吉布斯自由能最低最稳定,分别为-6.70和-9.02 kcal·moL^(-1)。利用ELISA试验进行亲和力测试,结果表明核酸适体S1和S2的亲和能力较强,S1:KD=67.02 nmol·L^(-1),R2=0.925 8,S2:KD=97.65 nmol·L^(-1),R2=0.795 1。核酸适体S1与过敏原Ses i 2的结合力和其他过敏原蛋白相比有显著差异,可视为具有特异性。本研究最终获得一条兼具良好亲和力和特异性的核酸适体S1,为芝麻过敏原快速检测提供了技术支撑。Sesame is one of the eight major food allergens.Rapid and accurate identification of allergens in sesame is important for preventing sesame allergies.Nucleic acid aptamers can effectively recognize target allergens and show promising application in allergen detection.To obtain specific nucleic acid aptamers for Ses i 2,the main allergen of sesame,Ses i 2 was used as the target.Ten rounds of screening with magnetic beads-based SELEX were conducted,and six candidate sequences(S1-S6)were obtained through highthroughput sequencing.Additionally,familial and homology analysis was performed for S1-S6,as well as secondary structure prediction.The results showed that the duplication rate of the six candidate nucleic acid aptamers reached 46.38%.The Gibbs free energies of these aptamers ranged from-9.02 kcal·mol^(-1) to-2.47 kcal·mol^(-1).According to the principle of free energy stability,the Gibbs free energies of S1 and S5 were the lowest,at-6.70 and-9.02 kcal·mol^(-1),respectively.Affinity tests using the ELISA assay showed that the nucleic acid aptamers S1 and S2 exhibited strong binding ability,with dissociation constants(KD)of 67.02 nmol·L^(-1),97.65 nmol·L^(-1),respectively.The R2 for S1 and S2 were 0.9258 and 0.7951,respectively.The binding of nucleic acid aptamer S1 to the allergen Ses i 2 was significantly different from that of other allergenic proteins,indicating specificity.In this study,a nucleic acid aptamer S1 possessing good affinity and specificity was successfully obtained,which provides technical support for the rapid detection of sesame allergens.

关 键 词：芝麻过敏原 Ses i 2 核酸适体 亲和力 

分 类 号：TS207.3[轻工技术与工程—食品科学]