Fontanel M L, Bazin H, Téoule R
CEA/Départment de Recherche Fondamentale sur la matière condensée/SESAM, Grenoble, France.
Nucleic Acids Res. 1994 Jun 11;22(11):2022-7. doi: 10.1093/nar/22.11.2022.
The ability of T4 polynucleotide kinase (PNK) to phosphorylate non-nucleosidic moieties 5'-attached to oligodeoxynucleotides (ODNs) has been investigated. Non-nucleosidic phosphoramidite units were prepared from ethane-1,2-diol and propane-1,3-diol backbones. Some of them corresponded to pure enantiomers. They were used to obtain the corresponding 5'-end modified oligothymidylates X(pdT)10. The free primary hydroxyl of the non-nucleosidic moieties (X) of these oligomers was phosphorylated by PNK. We report the stereoselective phosphorylation of the L form of the 5'-end attached non-nucleosidic chiral fragments; the non-chiral moieties were completely phosphorylated. Dimers of glycerol analogue and thymidine 3'-phosphate were not recognized by PNK and the shortest modified ODN able to be phosphorylated was a trinucleotide X(pdT)3. A modified X(pdT)10, bearing a cyclic abasic site (X) at its 5'-end, was prepared by chemical synthesis from 1,2-dideoxyribose phosphoramidite and was phosphorylated with a 90% yield.
对T4多核苷酸激酶(PNK)磷酸化与寡脱氧核苷酸(ODN)5'-相连的非核苷部分的能力进行了研究。非核苷亚磷酰胺单元由乙烷-1,2-二醇和丙烷-1,3-二醇主链制备。其中一些对应于纯对映体。它们被用于获得相应的5'-末端修饰的寡胸苷酸X(pdT)10。这些寡聚物的非核苷部分(X)的游离伯羟基被PNK磷酸化。我们报道了5'-末端连接的非核苷手性片段L型的立体选择性磷酸化;非手性部分被完全磷酸化。甘油类似物和胸苷3'-磷酸的二聚体不被PNK识别,能够被磷酸化的最短修饰ODN是三核苷酸X(pdT)3。通过化学合成由1,2-二脱氧核糖亚磷酰胺制备了一种在其5'-末端带有环状无碱基位点(X)的修饰X(pdT)10,并以90%的产率进行了磷酸化。