河豚毒素DNA适配子核心区域的识别  

Identification of Core Region of DNA Aptamers against Tetrodotoxin

作　　者：邵碧英[1] 陈文炳[1] 刘正才[1] 缪婷玉[1] 彭娟[1] 杨方[1] SHAO Biying;CHEN Wenbing;LIU Zhengcai;MIAO Tingyu;PENG Juan;YANG Fang(Fujian Provincial Key Laboratory of Inspection and Quarantine Technology Research,Fujian Entry-Exit Inspection and Quarantine Bureau, Fuzhou 350001, China)

机构地区：[1]福建出入境检验检疫局福建省检验检疫技术研究重点实验室,福建福州350001

出　　处：《食品科学》2017年第4期260-264,共5页Food Science

基　　金：国家质检总局科技计划项目(2014IK104)

摘　　要：用DNA folding form在线软件分析河豚毒素DNA适配子A3、G10的二级结构,根据二级结构,分别合成适配子A3、G10的各4个区域的5’端地高辛标记的序列。用丙烯酸-环氧乙烷珠子固定河豚毒素,测定抗地高辛碱性磷酸酶的适宜稀释倍数,用地高辛-抗地高辛碱性磷酸酶系统检测各区域与河豚毒素的亲和力,识别出适配子的核心区域。结果表明,抗地高辛碱性磷酸酶的适宜工作浓度为2×104倍稀释;合成的适配子A3的4个区域中,A3-2区域与河豚毒素的亲和力最强,即适配子A3的核心区域为A3-2区域,实际上含有47个核苷酸;合成的适配子G10的4个区域中,G10-4区域与河豚毒素的亲和力最强,即适配子G10的核心区域为G10-4区域,实际上含有56个核苷酸。The secondary structures of DNA aptamers A3and G10against tetrodotoxin were analyzed using DNA foldingform online software.Further,digoxin labeled5’end sequences of four regions in each aptamer were separately synthesized.Tetrodotoxin was fixed by acrylic acid-epoxy ethane beads.The core regions of these aptamers were identified usingdigoxin-anti digoxin alkaline phosphatase system to detect the affinity of these various regions with tetrodotoxin,afterdetermining the appropriate dilution ratio of anti-digoxin alkaline phosphatase.The results showed that the suitable workingconcentration of anti-digoxin alkaline phosphatase was2×104times dilution.The core region of aptamer A3was A3-2areawhich had the strongest affinity with tetrodotoxin among the four regions,including47nucleotides,while the core region ofaptamer G10was G10-4area,including56nucleotides.

关 键 词：河豚毒素 适配子 核心区域 亲和力 

分 类 号：Q78[生物学—分子生物学]