Gaynor James W, Bentley Joanne, Cosstick Richard
Department of Chemistry, University of Liverpool, Liverpool, L69 7ZD, UK.
Nat Protoc. 2007;2(12):3122-35. doi: 10.1038/nprot.2007.451.
Oligodeoxynucleotides containing 3'-S-phosphorothiolate (3'-PS) linkages have become useful tools for probing enzyme-catalyzed cleavage processes in DNA. This protocol describes the synthesis of the phosphorothioamidite monomers derived from thymidine and 2'-deoxycytidine, and their application to a fully automated procedure for synthesising oligodeoxynucleotides containing 3'-PS linkages. The synthesis of the 5'-protected-3'-amidites is achievable in 2 weeks with the DNA synthesis and purification taking another 1 week.
含有3'-硫代磷酸酯（3'-PS）键的寡脱氧核苷酸已成为探测DNA中酶催化切割过程的有用工具。本方案描述了源自胸苷和2'-脱氧胞苷的硫代磷酰胺单体的合成，以及它们在全自动合成含3'-PS键的寡脱氧核苷酸过程中的应用。5'-保护的3'-亚磷酰胺的合成可在2周内完成，DNA的合成和纯化再需要1周时间。
