Photoregulation of DNA polymerase I (Klenow) with caged fluorescent oligodeoxynucleotides.

Abstract:

The DNA polymerase reaction by Klenow fragment (KF) was efficiently regulated with UV light using a 25-mer caged fluorescent oligodeoxynucleotide (CFO) as the template. The CFO was functionalized with a fluorescein reporter (Fl) and photocleavable DABSYL quencher moiety (Dab). With Fl and Dab at adjacent cytidines in the middle at the template, KF was blocked from extending a complementary 12-mer primer. Upon UV photolysis of the DABSYL blocking group under aerobic conditions, fluorescein emission was restored and 50% of the primers were fully extended by KF.

Polymerases:

Topics:

Nucleotide Incorporation, Exonuclease Activity, Kinetic Parameters, Enzyme Substrate Interactions, Biotech Applications

Status:

new topics/pols set partial results complete validated

Results:

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