Complete template-directed enzymatic synthesis of a potential antisense DNA containing 42 methylphosphonodiester
M A Dineva1, S Chakurov, E K Bratovanova
1Laboratory of Biocatalysis, Bulgarian Academy of Sciences, Sofia.
Abstract:
P alpha-Methyl thymidine triphosphate was prepared through the pyrophosphorolysis of P alpha-methyl thymidine diphosphate P beta-diphenyl ester and tested as an alternative substrate for E. coli DNA polymerase 1 (Klenow fragment) using several template-primer systems requiring the formation of 1 to 42 methylphosphono diester bonds. The enzyme catalyzes the incorporation of a P-methyl thymidylic residue with (Sp)-configuration at a single site in a recessive 3'-end as well as at multiple sites along a growing 167 nucleotide long chain. The synthesis of a full length product, containing 42 sites of methylphosphonate incorporation was observed.
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