A homogeneous, high-throughput fluorescence resonance energy transfer-based DNA polymerase assay |
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| Authors: | Shapiro Adam Rivin Olga Gao Ning Hajec Laurel |
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| Institution: | Department of Biochemistry, Infection Discovery, AstraZeneca R&D Boston, Waltham, MA 02451, USA. adam.shapiro@astrazeneca.com |
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| Abstract: | A homogeneous, fluorescence resonance energy transfer (FRET)-based DNA polymerase assay that is suitable for high-throughput screening for inhibitors, and can also be used for steady-state kinetic investigations, is described. The activity, kinetic mechanism, and processivity of the isolated alpha subunit of DNA polymerase III, the product of the dnaE gene, from the gram-negative pathogen Haemophilus influenzae were investigated using the FRET assay. |
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| Keywords: | DNA polymerase Fluorescence resonance energy transfer FRET DnaE Pol III High-throughput |
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