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Nucleic Acids Research, Vol 26, Issue 9 2105-2111, Copyright © 1998 by Oxford University Press

ARTICLES

Synthesis and RNA polymerase incorporation of the degenerate ribonucleotide analogue rPTP

K Moriyama, K Negishi, MS Briggs, CL Smith, F Hill, MJ Churcher, DM Brown and D Loakes
Gene Research Centre, Okayama University, Tsushima, Okayama 700, Japan, Nycomed Amersham plc, Amersham Laboratories, White Lion Road, Amersham, Buckinghamshire HP7 9LL, UK.

The synthesis and enzymatic incorporation into RNA of the hydrogen bond degenerate nucleoside analogue 6-(beta-d-ribofuranosyl)-3, 4-dihydro-8H- pyrimido[4,5-c]-[1,2]oxazin-7-one (P) is described. The 5'-triphosphate of this analogue is readily incorporated by T3, T7 and SP6 RNA polymerases into RNA transcripts, being best incorporated in place of UTP, but also in place of CTP. When all the uridine residues in an HIV- 1 TAR RNA transcript are replaced by P the transcript has similar characteristics to the wild-type TAR RNA, as demonstrated by similar melting temperatures and CD spectra. The P-substituted TAR transcript binds to the Tat peptide ADP-1 with only 4-fold lowered efficiency compared with wild-type TAR.
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