Synthesis of symmetrical thiol-adenosine conjugate and 5′ thiol-RNA preparation by efficient one-step transcription
Synthesis of symmetrical thiol-adenosine conjugate and 5′ thiol-RNA preparation by efficient one-step transcription
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DOI：
10.1016/j.bmcl.2010.08.097
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发表时间：
2010-11-01
影响因子：
2.7
通讯作者：
Shi, Yongliang
中科院分区：
文献类型：
--
作者：
Huang, Faqing;Shi, Yongliang
化学合成了含有两个腺苷和缀合硫醇官能团（ThioAMP 二聚体）的对称转录起始子。在 T7 Phi 2.5 启动子下存在 ThioAMP 二聚体的情况下，转录可产生 5' 硫醇标记的 RNA (5' HS-RNA)，标记效率高达 90%，具体取决于 ThioAMP 二聚体与 ATP 的浓度比。所得 5' HS-RNA 可以直接使用，也可以在硫代丙基 Separose 6B 亲和柱纯化后使用。展示了 5' HS-RNA 的生物素化和金纳米颗粒-RNA 纳米复合物的形成。 (C) 2010 Elsevier Ltd. 保留所有权利。
A symmetrical transcription initiator containing two adenosines and conjugated thiol functionality (ThioAMP dimer) is chemically synthesized. Transcription in the presence of ThioAMP dimer under the T7 Phi 2.5 promoter yields 5' thiol-labeled RNA (5' HS-RNA) with up to 90% labeling efficiency, depending on the concentration ratio of ThioAMP dimer to ATP. The resulting 5' HS-RNA may be used directly or after thiopropyl Separose 6B affinity column purification. Biotinylation of 5' HS-RNA and formation of gold nanoparticle-RNA nanoplexes are demonstrated. (C) 2010 Elsevier Ltd. All rights reserved.