Wang Rui, Haruehanroengra Phensinee, Sheng Jia
Department of Chemistry and The RNA Institute, University at Albany, State University of New York, Albany, New York.
Curr Protoc Nucleic Acid Chem. 2017 Mar 2;68:4.72.1-4.72.13. doi: 10.1002/cpnc.22.
This unit describes the chemical synthesis of the S-geranyl-2-thiouridine (ges U) phosphoramidite and its incorporation into RNA oligonucleotides through solid-phase synthesis. Starting from the 2-thiouracil nucleobase and the protected ribose, the 2-thiouridine is synthesized and the geranyl functionality is introduced into the 2-thio position by using geranyl bromide as the geranylating reagent before the conversion of this modified nucleoside into a phosphoramidite building block. The modified phosphoramidite is used to make the geranyl-RNA oligonucleotides with a solid-phase DNA synthesizer. These RNA strands are then purified by ion-exchange HPLC before further structural and functional studies, such as base pairing and enzyme recognition, can be done. © 2017 by John Wiley & Sons, Inc.
本单元描述了S-香叶基-2-硫代尿苷（ges U）亚磷酰胺的化学合成及其通过固相合成掺入RNA寡核苷酸的过程。从2-硫代尿嘧啶碱基和受保护的核糖开始，合成2-硫代尿苷，并在将这种修饰的核苷转化为亚磷酰胺构建块之前，使用香叶基溴作为香叶基化试剂将香叶基官能团引入2-硫代位置。用固相DNA合成仪使用修饰的亚磷酰胺制备香叶基-RNA寡核苷酸。然后通过离子交换HPLC纯化这些RNA链，之后才能进行进一步的结构和功能研究，如碱基配对和酶识别。©2017约翰威立父子公司版权所有
