Hou Xiaorong, Wang Gang, Gaffney Barbara L, Jones Roger A
Rutgers, The State University of New Jersey, Piscataway, New Jersey.
Curr Protoc. 2023 Mar;3(3):e710. doi: 10.1002/cpz1.710.
This article describes procedures for preparation of deoxyguanosine and guanosine derivatives in which the guanine N contains a thiopropyl tether, protected as a tert-butyl disulfide. After incorporation into a DNA or RNA fragment, this tether allows site-specific cross-linking to a thiol of a protein or another nucleic acid. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation of diisopropyl-1-(tert-butylthio)-1,2-hydrazinedicarboxylate (4) Basic Protocol 2: Preparation of the 2'-deoxyguanosine N -propyl-tert-butyl disulfide phosphoramidite (12) Basic Protocol 3: Preparation of the guanosine N -propyl-tert-butyl disulfide phosphoramidite (20) Basic Protocol 4: Preparation of DNA fragments containing N -propyl-tert-butyl disulfide guanine Alternate Protocol: Preparation of RNA fragments containing N -propyl-tert-butyl disulfide guanine Basic Protocol 5: Conversion of N -propyl-tert-butyl disulfide to the free thiol, disulfide 5-thio-2-nitrobenzoic acid disulfide, or ethylamine disulfide.
本文介绍了脱氧鸟苷和鸟苷衍生物的制备方法，其中鸟嘌呤N含有一个硫代丙基连接基，以叔丁基二硫化物的形式进行保护。将其掺入DNA或RNA片段后，该连接基可实现与蛋白质或另一种核酸的硫醇进行位点特异性交联。© 2023作者。由Wiley Periodicals LLC出版的《当前实验方案》。基本方案1：二异丙基-1-（叔丁基硫代）-1,2-肼二羧酸酯（4）的制备 基本方案2：2'-脱氧鸟苷N-丙基-叔丁基二硫化物亚磷酰胺（12）的制备 基本方案3：鸟苷N-丙基-叔丁基二硫化物亚磷酰胺（20）的制备 基本方案4：含有N-丙基-叔丁基二硫化物鸟嘌呤的DNA片段的制备 替代方案：含有N-丙基-叔丁基二硫化物鸟嘌呤的RNA片段的制备 基本方案5：将N-丙基-叔丁基二硫化物转化为游离硫醇、二硫化物5-硫代-2-硝基苯甲酸二硫化物或乙胺二硫化物。
