不需要结构切换:用于小分子检测的可泛化外核酶介导的aptamer-based测试
Juan Canoura1, Zongwen Wang1,2, Haixiang Yu1
1Department of Chemistry and Biochemistry , Florida International University , 11200 Southwest Eighth Street , Miami , Florida 33199 , United States.
这项研究引入了一种新的检测小分子的方法, 目标结合保护体免受消化,使其能够在没有序列工程的情况下进行敏感,无标签的检测.
科学领域:
- 生物化学
- 分子生物学
- 分析化学
背景情况:
- 小分子与DNA结合可以改变其对外核酶消化的敏感性.
- 短DNA或RNA序列的aptamers用于分子识别和检测.
研究的目的:
- 开发一种敏感的,无标签的小分子检测方法,使用体和外核酶.
- 证明这种新型试验的广泛适用性和多重复合能力.
主要方法:
- 使用在目标结合时抵抗外核酶III消化的预折叠体.
- 使用双外核酶系统 (外核酶III和外核酶I) 进行选择性产品生成.
- 使用SYBR黄金等光染料量化剩余的双链DNA产品.
主要成果:
- 目标结合抑制了外核酶III的消化,留下一个可检测的双链产物.
- 不与标结合的受体被降解,而与标结合的受体保持完整.
- 证明了对脱异三硫酸盐,可卡因和ATP的敏感检测,包括同时多重检测.
结论:
- 这种双外核酶胺测定为小分子检测提供了通用,敏感和无标签的平台.
- 该方法可适应各种适应物和向物,适用于生物矩阵.
- 用于广泛分析应用的多种信号报告策略的集成潜力.
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