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Small-Scale Plasma Membrane Preparation for the Analysis of Candida albicans Cdr1-mGFPHis
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Small-Scale Plasma Membrane Preparation for the Analysis of Candida albicans Cdr1-mGFPHis

Small-Scale Plasma Membrane Preparation for the Analysis of Candida albicans Cdr1-mGFPHis

DOI:
10.3791/62592-v

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09:44 min

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June 13, 2021

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, , , , ,
1Sir John Walsh Research Institute, Faculty of Dentistry,University of Otago, 2Department of Microbiology, Faculty of Medicine,Chulalongkorn University, 3School of Biological Sciences,University of Auckland

Chapters

  • 00:04Introduction
  • 00:57Growing and Harvesting Yeast Cells
  • 02:26Isolation of Plasma Membranes
  • 04:14Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE)
  • 06:22Results: Analysis of Cdr1-mGFPHis Isolated from Plasma Membranes of Saccharomyces cerevisiae
  • 09:06Conclusion

Summary

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This article presents a small-scale plasma membrane isolation protocol for the characterization of Candida albicans ABC (ATP-binding cassette) protein Cdr1, overexpressed in Saccharomyces cerevisiae. A protease-cleavable C-terminal mGFPHis double tag with a 16-residue linker between Cdr1 and the tag was designed to facilitate the purification and detergent-screening of Cdr1.

Tags

Keywords: Plasma MembraneCandida AlbicansCdr1-mGFPHisMembrane ProteinSmall-scale IsolationATPaseDetergentCell HarvestingPre-cultureCell LysisCentrifugationHomogenization Buffer

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