Solid phase synthesis of 5′-diphosphorylated oligoribonucleotides and their conversion to capped m7 Gppp-oligoribonucleotides for U.S. as primers for influenza A virus RNA polymerase in vitro

G. G. Brownlee, E. Fodor, D. C. Pritlove, K. G. Gould, J. J. Dalluge

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Abstract

We have synthesized four different 5′-diphosphorylated oligoribonucleotides, varying in length from 11 to 13 nucleotides by a new solid phase method. After deprotectlon and partial purification the 5′-diphosphorylated oligoribonucleotides could be converted to capped (m7Gppp)-oligoribonucleotides using guanylyl transferase. Radlolabelled 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 ollgodeoxyribonucleotides and be capable of automation.

Original languageEnglish (US)
Pages (from-to)2641-2647
Number of pages7
JournalNucleic acids research
Volume23
Issue number14
DOIs
StatePublished - Jul 25 1995

Bibliographical note

Funding Information:
We thank Prof. G. Lowe for originally suggesting the phosphorylation method and for advice and Dr T. Claridge for the3IP NMR analysis. We acknowledge the encouragement of Drs M. Gait and B. Sproat This work was supported by an MRC project grant to GGB and an NIH grant GM29812 to Prof. J. A. McCloskey (University of Utah).

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