SUGAR-MODIFIED NUCLEOTIDE ANALOGS IN DNA-SYNTHESIS IN-VITRO

The substrate properties of dNTP analogs modified in the su,oar moiety were assessed in a cell-free system for DNA synthesis driven by Esche richia coli DNA polymerase Klenow fragment, reverse transcriptases of AMV, HIV-1, and M-MLV, and Thermus aquaticus DNA polymerase. Taq polym erase incorporated none of the compounds tested, whereas reverse trans criptases could incorporate all of them. The most efficient terminatio n substrate was deoxy-beta-D-glyceropent-3'-enofuranosyl)adenosine 5'- triphosphate; the least efficient was -deoxy-3'-fluoro-beta-D-arabinof uranosyl)adenosine 5'-triphosphate; 2'-azido-2',3'-didehydro-3'-deoxyt hymidine 5'-triphosphate, '-dideoxy-3'-fluoro-beta-D-ribofuranosyl)ade nosine 5'-triphosphate, and ideoxy-3'-fluoro-beta-D-arabinofuranosyl)a denosine 5'-triphosphate were intermediate.