Pieles U, Englisch U
Abteilung Chemie, Max-Planck-Institut für Experimentelle Medizin, Göttingen, FRG.
Nucleic Acids Res. 1989 Jan 11;17(1):285-99. doi: 10.1093/nar/17.1.285.
The psoralen derivative 4,5',8-trimethylpsoralen was covalently linked to the 5'-terminus of an 18mer oligodeoxyribonucleotide in the course of solid phase synthesis using phosphoroamidite chemistry. The derivative was introduced as a phosphitylation compound in the last cycle of the oligomer synthesis. The reagent was prepared by 4'-chloromethylation of 4,5',8-trimethylpsoralen, introduction of a linker by ethanediol and phosphitylation with chloro-[(beta-cyanoethoxy)-N,N-diisopropylamino]-phosphine. After oxydation and deprotection the 5'-psoralen modified oligodeoxyribonucleotide was characterised by HPLC. Hybridisation of the psoralen-modified oligomer to a complementary single stranded 21mer followed by irradiation at 350 nm revealed a photo-cross-linked double-stranded DNA fragment analysed on denaturing polyacrylamide gels. The cross-link could be reversed upon irradiation at 254nm.
在使用亚磷酰胺化学的固相合成过程中，补骨脂素衍生物4,5',8-三甲基补骨脂素与一个18聚体寡脱氧核糖核苷酸的5'-末端共价连接。该衍生物在寡聚物合成的最后一个循环中作为磷酰化化合物引入。该试剂通过4,5',8-三甲基补骨脂素的4'-氯甲基化、用乙二醇引入连接子以及用氯-[(β-氰基乙氧基)-N,N-二异丙基氨基]-膦进行磷酰化来制备。氧化和脱保护后，5'-补骨脂素修饰的寡脱氧核糖核苷酸通过高效液相色谱进行表征。补骨脂素修饰的寡聚物与互补的单链21聚体杂交，然后在350nm下照射，得到一个在变性聚丙烯酰胺凝胶上分析的光交联双链DNA片段。在254nm照射下，交联可以逆转。
