优化的合成和纯化腺苷-叶酸缀合物的方法，用作制备修饰 RNA 中的转录起始物。
Optimized method for the synthesis and purification of adenosine--folic acid conjugates for use as transcription initiators in the preparation of modified RNA.
机构信息
Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN 47907, USA.
出版信息
Methods. 2011 Jun;54(2):260-6. doi: 10.1016/j.ymeth.2010.12.007. Epub 2010 Dec 14.
Abstract
We present an optimized synthetic strategy for the attachment of molecules to 5'-adenosine monophosphate (AMP), which can then be used to label the 5'-end of RNA by T7 RNA polymerase mediated in vitro transcription. Through the use of a boronate affinity gel, we have developed an efficient route to the preparation of folate conjugated AMP with high yields and purity. Affi-Gel boronate is an affinity resin that selectively binds nucleoside and nucleoside derivatives at pH>7.5 and releases them at pH<6.5. This resin is used to efficiently bind and purify ribonucleotides such as AMP. This allows for the addition of a large excess of reactants and reagents in order to drive the reaction to completion and then allow easy purification without HPLC. The synthesis can be successfully scaled up to produce large quantities of AMP conjugates.
摘要
我们提出了一种优化的合成策略，用于将分子连接到 5'-腺苷一磷酸 (AMP) 上，然后可以通过 T7 RNA 聚合酶介导的体外转录将其用于标记 RNA 的 5'-末端。通过使用硼酸亲和凝胶，我们开发了一种高效的方法来制备具有高产率和高纯度的叶酸偶联 AMP。Affi-Gel 硼酸亲和树脂是一种亲和树脂，在 pH>7.5 时选择性地结合核苷和核苷衍生物，并在 pH<6.5 时将其释放。该树脂用于有效结合和纯化 AMP 等核糖核苷酸。这允许添加大量过量的反应物和试剂，以推动反应完成，然后允许在没有 HPLC 的情况下轻松纯化。该合成可以成功放大以生产大量的 AMP 缀合物。
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