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A G-quadruplex DNA-affinity Approach for Purification of Enzymatically Active G4 Resolvase1

Article DOI: 10.3791/55496-v • 11:25 min • March 18th, 2017
March 18th, 2017 •
*1, *2, 3, 4, 1, 3
1Department of Cancer Biology, Wake Forest School of Medicine, 2YX Genomics, 3Department of Biology, Ball State University, 4Department of Hematology and Oncology, Roper St. Francis Hospital
* These authors contributed equally

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요약
Automatic Translation

G4 Resolvase1 binds to G-quadruplex (G4) structures with the tightest reported affinity for a G4-binding protein and represents the majority of the G4-DNA unwinding activity in HeLa cells. We describe a novel protocol that harnesses the affinity and ATP-dependent unwinding activity of G4-Resolvase1 to specifically purify catalytically active recombinant G4R1.

Tags

G-quadruplex DNA-affinity Purification Enzymatically Active G4 Resolvase1 Protein Isolation ATP-dependent Purification Catalytically Active Enzyme Recombinant Protein In Vitro Biochemical Assays Misfolded Enzyme ATP Elution G-quadruplex Helicase Bacterial Expression System TN Buffer Lysozyme Digestion
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