New affinity resin for purification of cap-binding proteins.
New affinity resin for purification of cap-binding proteins.
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用于纯化帽结合蛋白的新型亲和树脂。
DOI：
10.1081/ncn-200061782
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Darzynkiewicz,Edward
中科院分区：
文献类型：
--
作者：
Jankowska-Anyszka,Marzena;Nogalski,Maciej;Darzynkiewicz,Edward
帽结合蛋白可识别 RNA 聚合酶 II 转录物 5' 末端的帽结构，已使用附有单核苷酸帽类似物的亲和树脂进行常规分离和纯化。在这里，我们提出了一种新方法，其中二核苷酸帽类似物 m7GpppG 已与 EAH-Sepharose 连接。该方法基于帽结构内第二个核苷酸的2', 3'-顺式二醇与乙酰丙酸的衍生化，以及随后通过其羧基将所得缩醛与氨基己基琼脂糖偶联。
Cap binding proteins, which recognize the cap structure present at 5′ termini of RNA polymerase II transcripts, have been routinely isolated and purified using affinity resins with mononucleotide cap analogs attached. Here we present a new methodology in which dinucleotide cap analog, m7GpppG, has been linked to the EAH-Sepharose. The method is based on derivatization of 2′, 3′-cis diol of the second nucleotide within the cap structure, with levulinic acid, and subsequent coupling of resulted acetal through its carboxylic group with aminohexyl-agarose.