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Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae
JoVE Journal
Biology
This content is Free Access.
JoVE Journal Biology
Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae
DOI:

15:12 min

May 10, 2014

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Chapters

  • 00:05Title
  • 01:46Solubilization of Microsomes
  • 03:49Nickel-affinity Purification of CFTR
  • 05:33Gel Permeation Chromatography (GPC) Purification of CFTR
  • 06:57Reconstitution of CFTR
  • 07:51Measurement of ATPase Activity
  • 09:35Results: Isolation of Milligram Quantities of CFTR by Two Methods
  • 14:28Conclusion

Summary

Automatic Translation

Heterologous expression and purification of the cystic fibrosis transmembrane conductance regulator (CFTR) are significant challenges and limiting factors in the development of drug therapies for cystic fibrosis. This protocol describes two methods for the isolation of milligram quantities of CFTR suitable for functional and structural studies. 

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