Abstract
Acyclic analogues of oligothymidylate and oligoadenylate and their alternating copolymers were synthesized to study their thermal melting, their stability against snake venom phosphodiesterase and their primer/template properties using the Klenow fragment of the Escherichia coli DNA polymerase I enzyme. Acyclic dodecaadenylate (GlyA) 12 hybridized to dodecathymidylate p(dT) 12, and the complex presented a sharp melting with a T m at 24°C. This association was confirmed by circular dichroism curves which were similar to those of the natural oligonucleotide duplexes in A-conformation. (GlyA) 12 proved very stable against snake venom phosphodiesterase hydrolysis. The reaction rate was more than 10000 times slower than that of p(dT) 12. (GlyA) 12 served as a primer for the Klenow DNA polymerase. When (GlyA) 12 was complexed with the poly(dT) template, the enzyme polymerized dATP but the reaction was much slower than with the (GlyT) 12 primer. Molecular modelling of atactic (GlyA) 12·(dT) 12 of the A-conformation indicates that this conformation is energetically possible.
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