Synthesis and enzymatic incorporation of a novel, bicyclic pyrimidine nucleoside: a thymidine mimic

✍ Scribed by David Loakes; Daniel M Brown; Stephen A Salisbury; Mark G McDougall; Constantin Neagu; Satyam Nampalli; Shiv Kumar

• Publisher: Elsevier Science
• Year: 2003
• Tongue: French
• Weight: 170 KB
• Volume: 44
• Category: Article
• ISSN: 0040-4039
• DOI: 10.1016/s0040-4039(03)00567-7

✦ Synopsis

Nucleophilic ring-opening and rearrangement reaction of a furanopyrimidine nucleoside with anhydrous hydrazine provided a novel, 6,6-bicyclic pyrimidopyridazin-7-one nucleoside (dH, 4), whose structure was confirmed by X-ray crystallography. This novel nucleoside was converted to its 5%-triphosphate (dHTP) for studies with DNA polymerases and incorporated into a template by using standard phosphoramidite chemistry. In the template, dH directed the incorporation of dATP and to a lesser extent dGTP into the transcript and dHTP was efficiently incorporated at the 3%-end of a primer opposite dA using both exonuclease free Klenow fragment (KF exo-) and Taq DNA polymerases and extended with natural dNTPs.