赭曲霉毒素A核酸适配体筛选.pdf

第6卷第9期
食品安全质量检测学报
Vol 6 No 9
2015年9月
Journal of Food Salcly and Quality
Scp.,2015
赭曲霉毒素A核酸适配体筛选
刘继红123,胡颖4,洪慧杰23,张玲,2,3,王红旗123
(1.河南省农业科学院农业质量标准与检測技术研究所,郑州450002.河南省粮食质量安全与检测重点实验室,郑州
450002:3.农业部农产品质量安全风险评估实验室(郑州),郑州4500024.河南农业大学,郑州45000)
摘要:目的筛选小麦和其他相关食品中常见的赭曲霉毒素A真菌毒素核酸适配体。方法采用固态靶标
筛选策略和磁分离的筛选方法筛选赭曲霉毒素A的核酸适配体。结果获得的赭曲霉毒素A核酸适配体亲和
力较高、解离常数为纳摩尔级:特异性好、与赭曲霉毒素A结构相似性化合物如赭曲霉毒素B、N-乙酰苯丙氨
酸或华法令不结合或结合力非常弱。结论本文采用固态靶标筛选策略和磁分离的SELEⅩ筛选方法获得了赭
曲霉毒素A的高特异性、高亲和力核酸适配体、有望在农产品质量安全领域替代传统抗体开发真菌毒素快速
检测传感器或制备固相亲和柱
关键词:真菌毒素;赭曲霉毒素,核酸适配体;指数富集的配基系统进化技术;固相亲和柱
Screening of DNA aptamer of ochratoxin A
LIU Ji-hong, HU Ying* HONG Hui-jie., ZHANG Ling.2, WANG Hong-qi.3
(1. Nstitute fairy Standad and esting echnolo for 4 -rodae, denan Academy ofdgricu u a/Science
Zhen zho 450002, China: 2. Heman Key I boratory f(in ality and Safety and Testing, Zhengzhou 450002, hina
3. Laboratory o/Ouality Safely Risk ssessmmem 0r4g70-procacts(Zhengzhou), Ministry o/dgriczlure
Zhengzhou 450002, China; 4. Henan griculnral University, Zhengzhou 450002, China)
ABSTRACT: Objective To screen aptamer of ochratoxin A(OTA)in wheat and other related food
Methods The solid-tarect sclcction stratcgy and magnctic-scparation sclcction mcthod were adoptcd to scrccn
aptamer of ochratoxin A. Results The OIA aptamer had high affinity with nanomolar dissociation constants
and good specificity non-reco nizing, and it had weak affinity towards structure similarity compounds of OTA
such as Ochratoxin B(O] B), N-acelyl-i-phenylalanine and warfarin. Conclusion The OIA aptamer that go
Irom solid-targel seleclion strategy and magnelic-separalion selection method, has high allinity and good
specificity. Instead of traditional antibody, the OTA aptamer may be used to develop sensor and allinity
chromatography column for mycotoxins rapid detection in the ficld of agricultural products
KEY WORDS: mycotoxin ochratoxin aptamers, systematic cvolution of ligands by exponential cnrichment
affinity column
基金项目:国家公益性行业(农业)项目(201203094),国家自然科学基金项目(21305031),农业部重点项目(201-G5)
Fund: Supported by Spccial Fund for Agro-scicntific Rcscarch in the Public Intcrcst(201203094) the National Natural Scicncc Foundation of
China(21305031)and the Key Project of the Ministry of Agriculture (2011-(
通讯作者:王红旗,副研究员,主要研究方向为农产品质量安全检测技术。T-mal:huda2000c0126.com
Honan Academy of Agricultural Seicnce No 116, Huayuan Road, Jinshui Distict h n zhou 450002, China. E-mail: huda2000 126 s
Corresponding author: WANG Hong-qi, Associate Rcscarchcr, Institutc of Quality Standard and Testing Tcchnology for Agro-produ