Aptamer beacons for the direct detection of proteins
Aptamer beacons for the direct detection of proteins
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DOI：
10.1006/abio.2001.5169
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发表时间：
2001-07-15
影响因子：
2.9
通讯作者：
Stanton, M
中科院分区：
文献类型：
--
作者：
Hamaguchi, N;Ellington, A;Stanton, M
我们设计了一类新的分子，我们称之为适配子信标，用于检测广泛的配体。与分子信标类似，适配子信标可以采用两种或两种以上的构象，其中一种允许配体结合。荧光猝灭对用于报告配体结合引起的构象变化。将抗凝血酶适配子设计成适配子信标，在5‘端添加核苷酸，这些核苷酸与适配子3’端的核苷酸互补。在没有凝血酶的情况下，添加的核苷酸将与3‘端形成双链，迫使适体信标进入茎环结构。在凝血酶存在的情况下，适体信标形成配体结合结构。这种构象变化导致连接到5‘端的荧光团和连接到3’端的猝灭剂之间的距离改变。适配子信标可以作为检测蛋白质和其他化合物的灵敏工具。(C)2001年学术出版社。
We have designed a new class of molecules, which we term aptamer beacons, for detecting a wide range of ligands. Similar to molecular beacons, aptamer beacons can adopt two or more conformations, one of which allows ligand binding. A fluorescence-quenching pair is used to report changes in conformation induced by ligand binding. An anti-thrombin aptamer was engineered into an aptamer beacon by adding nucleotides to the 5 ' -end which are complementary to nucleotides at the 3 ' -end of the aptamer. In the absence of thrombin, the added nucleotides will form a duplex with the 3 ' -end, forcing the aptamer beacon into a stem-loop structure. In the presence of thrombin, the aptamer beacon forms the ligand-binding structure. This conformational change causesa change in the distance between a fluorophore attached to the 5 ' -end and a quencher attached to the 3 ' -end. Aptamer beacon can be a sensitive tool for detecting proteins and other chemical compounds. (C) 2001 Academic Press.