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Nucleic Acids Research, 1995, Vol. 23, No. 14 2641-2647
© 1995

RNA

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

G.G. Brownlee*, E. Fodor, D.C. Pritlove, K.G. Gould and J.J. Dalluge1

Chemical Pathology Unit, Sir William Dunn School of Pathology, University of Oxford South Parks Road, Oxford OX1 3RE, UK 1Department of Biochemistry, University of Utah Salt Lake City, UT 84132, USA

*To whom correspondence should be addressed

Received April 15, 1995. Accepted June 12, 1995.

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.


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