Solid phase synthesis of 5'-diphosphorylated oligoribonucleotides and their conversion to capped m7Gppp-oligoribonucleotides for use as primers for influenza A virus RNA polymerase in vitro.
We have synthesized four different 5'-diphosphorylated oligoribonucleotides, varying in length from 11 to 13 nucleotides by a new solid phase method. After deprotection and partial purification the 5'-diphosphorylated oligoribonucleotides could be converted to capped (m7Gppp) oligoribonucleotides using guanylyl transferase. Radiolabelled capped oligoribonucleotides acted as primers for the influenza A virus RNA polymerase in vitro. The solid phase method described here should also allow the addition of 5'-diphosphates to synthetic oligodeoxyribonucleotides and be capable automation.
Brownlee
G G
GG
Chemical Pathology Unit, Sir William Dunn School of Pathology, University of Oxford, UK.
Fodor
E
E
Pritlove
D C
DC
Gould
K G
KG
Dalluge
J J
JJ
D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D009980:Influenza A virus / Q000201:enzymology* / Q000235:genetics*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009843:Oligoribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D010766:Phosphorylation; D012313:RNA / Q000138:chemical synthesis* / Q000737:chemistry; D012315:RNA Caps / Q000138:chemical synthesis* / Q000737:chemistry; D012367:RNA, Viral / Q000235:genetics
D009843:Oligoribonucleotides; D012315:RNA Caps; C034477:RNA primers; D012367:RNA, Viral; D012313:RNA; D012321:DNA-Directed RNA Polymerases