Abstract:
:The ability of several DNA polymerases to catalyze the template-directed synthesis of duplex oligonucleotides containing a base pair between a nucleotide with anhydrohexitol ring and its natural complement has been investigated. All DNA polymerases were able to accept the chemically synthesized anhydrohexitol triphosphate as substrate and to catalyze the incorporation of one anhydrohexitol nucleotide. However, only family B DNA polymerases succeeded in elongating the primer after the incorporation of an anhydrohexitol nucleotide. In this family, Vent (exo(-)) DNA polymerase is the most successful one and was therefore selected for further investigation. Results revealed that at high enzyme concentrations six hATPs could be incorporated; however, a selective incorporation proved only feasible under experimental conditions where no more than two analogues could be inserted. Also the synthesis of a mixed HNA-DNA sequence was examined. Kinetic parameters for incorporation of one anhydrohexitol adenine nucleoside were similar to those of its natural analogue.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Vastmans K,Pochet S,Peys A,Kerremans L,Van Aerschot A,Hendrix C,Marlière P,Herdewijn Pdoi
10.1021/bi001297gsubject
Has Abstractpub_date
2000-10-24 00:00:00pages
12757-65issue
42eissn
0006-2960issn
1520-4995pii
bi001297gjournal_volume
39pub_type
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