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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

Article DOI: 10.3791/62592-v • 09:44 min • June 13th, 2021
June 13th, 2021 •
*1, *1, 1, 1,2, 3, 1
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
* These authors contributed equally

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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.

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Small-scale Plasma Membrane Preparation Analysis Candida Albicans Cdr1-mGFPHis Economical Fast Reliable Easy Method Characterize Membrane Proteins Eukaryotic Model Organism Saccharomyces Cerevisiae Technology Membrane Protein Expression Levels ATP's Activities Detergent Purification User-friendly OD 600 Mitochondrial Contamination ATPase Activities Pre-culture YPD Medium Incubate Cell Density Optical Density Units (ODU) Logarithmic Face Cells Centrifuging Wash Cells Ice Cold Sterile Double Distilled Water
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