Seelig B, Jäschke A
Institut für Biochemie, Freie Universität Berlin, Thielallee 63, D-14195 Berlin.
Bioconjug Chem. 1999 May-Jun;10(3):371-8. doi: 10.1021/bc980085h.
We give a detailed account on the enzymatic synthesis of RNA conjugates by T7 RNA polymerase using modified initiator nucleotides during transcription. Following two different routes, ternary conjugates of guanosine-5'-monophosphate, poly(ethylene glycol), and anthracene were synthesized via phosphoramidite intermediates and characterized by a variety of spectroscopic techniques. Up to a degree of polymerization nPEG of about 17, these conjugates were efficiently incorporated into RNA by T7 RNA polymerase at the 5'-termini, thereby giving access to RNA conjugates required for biochemical studies as well as for the exploration of the catalytic potential of ribonucleic acids. The resulting conjugates are intact and functional.
我们详细阐述了在转录过程中,T7 RNA聚合酶使用修饰的起始核苷酸进行RNA缀合物的酶促合成。通过两条不同的路线,经由亚磷酰胺中间体合成了鸟苷-5'-单磷酸、聚乙二醇和蒽的三元缀合物,并通过多种光谱技术对其进行了表征。对于聚合度nPEG约为17的情况,这些缀合物能被T7 RNA聚合酶有效地掺入到5'-末端的RNA中,从而获得生物化学研究以及探索核糖核酸催化潜力所需的RNA缀合物。所得的缀合物完整且具有功能。