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  • Rapid method for chromatin immunoprecipitation (ChIP) assay in a dimorphic fungus, Candida albicans.

Rapid method for chromatin immunoprecipitation (ChIP) assay in a dimorphic fungus, Candida albicans.

Journal of microbiology (Seoul, Korea) (2019-06-13)
Jueun Kim, Jung-Shin Lee
ABSTRACT

A chromatin immunoprecipitation (ChIP) assay is a method to identify how much a protein of interest binds to the DNA region. This method is indispensable to study the mechanisms of how the transcription factors or chromatin modifications regulate the gene expression. Candida albicans is a dimorphic pathogenic fungus, which can change its morphology very rapidly from yeast to hypha in response to the environmental signal. The morphological change of C. albicans is one of the critical factors for its virulence. Therefore, it is necessary to understand how to regulate the expression of genes for C. albicans to change its morphology. One of the essential methods for us to understand this regulation is a ChIP assay. There have been many efforts to optimize the protocol to lower the background signal and to analyze the results accurately because a ChIP assay can provide very different results even with slight differences in the experimental procedure. We have optimized the rapid and efficient ChIP protocol so that it could be applied equally for both yeast and hyphal forms of C. albicans. Our method in this protocol is also comparatively rapid to the method widely used. In this protocol, we described our rapid method for the ChIP assay in C. albicans in detail.

MATERIALS
Product Number
Brand
Product Description

Supelco
Ethanol, absolute for analysis EMSURE® ACS,ISO,Reag. Ph Eur
Sigma-Aldrich
Proteinase K from Tritirachium album, buffered aqueous glycerol solution, for molecular biology, ≥800 units/mL