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

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9.5K Views

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11:25 min

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March 18th, 2017

DOI :

10.3791/55496-v

March 18th, 2017

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Eric D. Routh*¹, Steven D. Creacy*², Peter E. Beerbower³, Steven A. Akman⁴, James P. Vaughn¹, Philip J. Smaldino³

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

Podsumowanie

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Keywords G quadruplex DNA

G4 Resolvase1

Enzymatic Purification

ATP dependent Purification

Recombinant Protein

Bacterial Expression

Catalytically Active Enzyme

Streptavidin Paramagnetic Beads

Rozdziały w tym wideo

0:05

Title

0:54

Lyse Bacterial Pellets Expressing Human Recombinant G4 Resolvase1 (rG4R1)

3:06

Prepare G4-magnetic Beads

4:07

Bind Histidine-tagged rG4R1 to Cobalt Beads

5:47

Elute rG4R1 from Cobalt Beads

7:07

Bind rG4R1 to G4-bound SPB and Elute in an ATP-dependent Manner

8:55

Results: Analysis and Quantification of Catalytically Active, Purified rG4R1

10:06

Conclusion

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