Add time:09/02/2019         Source:sciencedirect.com

In the present study, templates containing a specific segment of the Gγ-globin gene were constructed and incorporation of 3′-azido-3′-deoxythymidine 5′-triphosphate (AZT-TP) or 2′,3′-dideoxythymidine 5′-triphosphate (ddTTP) into these templates was compared to that observed in M13 bacteriophage DNA. Investigations on the intrinsic fidelity of T7 DNA polymerase in reactions with AZT-TP and without deoxythymidine 5′-triphosphate, resulted in DNA synthesis beyond the first T site, suggesting that other normal deoxynucleotides misincorporated at these T sites. Modified T7 DNA polymerase incorporated AZT-TP into T sites of elongating DNA strands. Chain termination at noncomplementary sites was also observed with AZT-TP when a genomic DNA template was used and interestingly, this phenomenon was not detected in the presence of a M13 DNA template. These DNA template-dependent effects were not detected with either ddTTP, 2′,3′-dideoxycytidine 5′-triphosphate, or 2′,3′-didehydro-2′,3′-dideoxythymidine 5′-triphosphate (D4T-TP). A variation in the extent of chain termination at T sites was observed with D4T-TP suggesting that each 2′,3′-dideoxynucleoside may exhibit unique chain termination patterns along with the template sequence.

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