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Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles
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Biochemistry
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JoVE Journal Biochemistry
Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles

Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles

DOI:
10.3791/60009-v

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13:16 min

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December 31, 2019

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, ,
1Department of Biological Sciences,Bowling Green State University

Chapters

  • 00:05Title
  • 01:18Generation of Inside-out Membrane Vesicles
  • 04:21Western Blot and Orientation of Transporter Assay
  • 06:46Transport Assay
  • 08:44Results: Characterization of Membrane Transporters
  • 11:05Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a method for the characterization of proton-driven membrane transporters in membrane vesicle preparations produced by heterologous expression in E. coli and lysis of cells using a French press.

Tags

Keywords: Membrane TransportersHeterologous ExpressionE. ColiMembrane VesiclesProtein GradientsSubstrate TransportAffinityCompetition AssaysSubstrate ActivationFrench PressUltracentrifugation

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