Sinha N D, Cook R M
Chemistry Division, Biosearch Inc., San Rafael, CA 94901.
Nucleic Acids Res. 1988 Mar 25;16(6):2659-69. doi: 10.1093/nar/16.6.2659.
Syntheses of H-phosphonate salts (4a-e) of N/S-protected alcohols such as 6-aminohexan-1-ol, 3-mercaptopropan-1-ol and 6-mercaptohexan-1-ol are described using 2-chloro-5,6-benzo-1,3,2-phosphorin-4-one (2) as the phosphonylating agent. The H-phosphonate salts (4a-e), in the presence of pivaloyl chloride or adamantoyl chloride as an activator, were coupled to the 5'-end of synthetic oligonucleotides on solid supports to produce amino or thio-linked oligonucleotides. Following deprotection and purification, fluorescent dyes, biotin derivatives and poly-L-lysine-maleimide were separately attached to the functionalised oligonucleotides. Identical derivatized oligomers were obtained with cyanoethyl-N,N-diisopropylamidite chemistry and amidites (5a-e) of the respective alcohols.
描述了使用2-氯-5,6-苯并-1,3,2-磷杂环丁烷-4-酮(2)作为膦酰化剂合成N/S保护醇(如6-氨基己醇、3-巯基丙醇和6-巯基己醇)的H-膦酸盐(4a - e)。在新戊酰氯或金刚烷酰氯作为活化剂存在的情况下,将H-膦酸盐(4a - e)与固相支持物上合成寡核苷酸的5'-末端偶联,以产生氨基或硫连接的寡核苷酸。脱保护和纯化后,将荧光染料、生物素衍生物和聚-L-赖氨酸-马来酰亚胺分别连接到功能化的寡核苷酸上。使用氰基乙基-N,N-二异丙基亚磷酰胺化学方法和相应醇的亚磷酰胺(5a - e)获得了相同的衍生寡聚物。