Enzymatic and NMR analysis of oligoribonucleotides synthesized with 2'-tert-butyldimethylsilyl protected cyanoethylphosphoramidite monomers.
The regioisomeric integrity of the internucleotide phosphate linkage in synthetic RNA using 2'-tert-butyldimethylsilyl protection was examined using enzymatic and NMR techniques. Two sets of DNA-RNA hybrid nonamers, T3XT5 and T5XT3 (where X = rA, rC, rG and U) and the tetramer AGCU were analyzed. Enzyme catalyzed hydrolysis of the nonamers with ribonuclease T2 showed that the linkage at the ribonucleotide was the desired 3'-5'. A control nonamer with a 2'-5' linkage was subjected to the enzyme, and showed no cleavage. High-resolution proton NMR of the tetramer also gave a favorable comparison with the same molecule obtained by non-chemical means.
Wang
Y Y
YY
Department of Chemistry, Syracuse University, NY 13244.
Lyttle
M H
MH
Borer
P N
PN
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004722:Endoribonucleases / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D009843:Oligoribonucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D017646:Organosilicon Compounds; D012825:Silicon
D009843:Oligoribonucleotides; D017646:Organosilicon Compounds; C009660:t-butyldimethylsilyl compounds; D004722:Endoribonucleases; C022543:ribonuclease T(2); D012825:Silicon