Book chapter
Conditional Protein Depletion in the Analysis of Antifungal Drug Resistance in Candida glabrata
Antifungal Drug Resistance, Vol.2658, pp.191-200
Methods in molecular biology (Clifton, N.J.), 2658, Humana
2023
DOI: 10.1007/978-1-0716-3155-3_13
PMID: 37024703
Abstract
This chapter illustrates a method to generate Candida glabrata conditional depletion mutants for SNF2, an ATPase subunit of the SWI/SNF chromatin remodeling complex with potential roles in the response to azole drugs. The strategy employed utilizes a plant-specific proteolysis pathway which allows for the rapid degradation of a target protein in the presence of the phytohormone, auxin. The steps taken to generate strains expressing the auxin-inducible plant F-box protein, Tir1, and in which the auxin-binding target, IAA17, is C-terminally fused to Snf2 are described. This acute depletion strategy is suitable for studying the effects of the loss of growth-critical proteins. The rapid depletion afforded by the auxin-induced degradation avoids the potential complications of a null allele causing a severe growth defect and allows a more rapid assessment of the consequences of reduced levels of a protein of interest.
Details
- Title: Subtitle
- Conditional Protein Depletion in the Analysis of Antifungal Drug Resistance in Candida glabrata
- Creators
- Thomas P Conway - University of IowaW Scott Moye-Rowley - Department of Molecular Physiology and Biophysics, Carver College of Medicine, University of Iowa, Iowa City, IA, USA. scott-moye-rowley@uiowa.edu
- Resource Type
- Book chapter
- Publication Details
- Antifungal Drug Resistance, Vol.2658, pp.191-200
- Publisher
- Humana; New York, NY
- Series
- Methods in molecular biology (Clifton, N.J.); 2658
- DOI
- 10.1007/978-1-0716-3155-3_13
- PMID
- 37024703
- ISSN
- 1064-3745
- eISSN
- 1940-6029
- Language
- English
- Date published
- 2023
- Academic Unit
- Molecular Physiology and Biophysics
- Record Identifier
- 9984385054002771
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