Preparation and purification of 5'-amino-5'-deoxyguanosine-5'-N-phosphoramidate and its initiation properties with T7 RNA polymerase.
5'-Amino-5'-deoxyguanosine-5'-N-phosphoramidate (GNHP) was synthesised in four steps from guanosine. Its identity was confirmed using spectroscopic and kinetic studies. A chromatographic method for the purification of this unstable compound was developed. GNHP was found to initiate T7 RNAP promoted transcriptions to afford 5'-O3P-NH-RNA, which hydrolyses readily to yield 5'-H2N-RNA that can be conjugated to activated esters.
Williamson
David
D
Department of Chemistry, University Science Laboratories, South Road, Durham, DH1 3LE, England. d.r.w.hodgson@durham.ac.uk.
Hodgson
David R W
DR
D002845:Chromatography; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification; D006151:Guanosine / Q000737:chemistry*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D014764:Viral Proteins / Q000737:chemistry*
C526213:5'-amino-5'-deoxyguanosine-5'-N-phosphoramidate; D014764:Viral Proteins; D006151:Guanosine; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D003849:Deoxyguanosine