Chapter title |
Conditional Protein Depletion in the Analysis of Antifungal Drug Resistance in Candida glabrata.
|
---|---|
Chapter number | 13 |
Book title |
Antifungal Drug Resistance
|
Published in |
Methods in molecular biology, January 2023
|
DOI | 10.1007/978-1-0716-3155-3_13 |
Pubmed ID | |
Book ISBNs |
978-1-07-163154-6, 978-1-07-163155-3
|
Authors |
Conway, Thomas P, Moye-Rowley, W Scott, Thomas P. Conway, W. Scott Moye-Rowley, Conway, Thomas P., Moye-Rowley, W. Scott |
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. |
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