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| Status |
Public on Nov 15, 2008 |
| Title |
A toolbox for epitope-tagging and genome-wide location analysis in Candida albicans |
| Organism |
Candida albicans |
| Experiment type |
Genome binding/occupancy profiling by array
|
| Summary |
Background : Candida albicans is a diploid pathogenic fungus not yet amenable to routine genetic investigations. Understanding aspects of the regulation of its biological functions and the assembly of its protein complexes would lead to further insight into the biology of this common disease-causing microbial agent.
Results: We have developed a toolbox allowing in vivo protein tagging by PCR-mediated homologous recombination with TAP, HA and MYC tags. The transformation cassettes were designed to accommodate a common set of integration primers. The tagged proteins can be used to perform tandem affinity purification (TAP) or chromatin immunoprecipitation coupled with microarray analysis (ChIP-CHIP). Tandem affinity purification of C. albicans Nop1 revealed the high conservation of the small processome composition in yeasts. Data obtained with in vivo TAP-tagged Tbf1, Cbf1 and Mcm1 recapitulates previously published genome-wide location profiling by ChIP-CHIP. We also designed a new reporter system for in vivo analysis of transcriptional activity of gene loci in C. albicans.
Conclusion: This toolbox provides a basic setup to perform purification of protein complexes and increase the number of annotated transcriptional regulators and genetic circuits in C. albicans.
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| Overall design |
Two independent biological replicates of ChIP-CHIP of Mcm1-TAP in yeast and hyphal states. ChIP-CHIP of Cbf1-TAP and Tbf1-TAP.
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| |
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| Contributor(s) |
Lavoie H, Sellam A, Askew C, Nantel A, Whiteway M |
| Citation(s) |
19055720 |
| |
| Submission date |
Oct 31, 2008 |
| Last update date |
Mar 20, 2012 |
| Contact name |
Hugo Lavoie |
| E-mail(s) |
hugo.lavoie@nrc-cnrc.gc.ca
|
| Phone |
514-496-6154
|
| Fax |
514-496-6213
|
| Organization name |
CNRC-IRB
|
| Department |
Genetics
|
| Street address |
6100 Royalmount
|
| City |
Montreal |
| State/province |
Quebec |
| ZIP/Postal code |
H4P 2R2 |
| Country |
Canada |
| |
|
| Platforms (1) |
| GPL6474 |
Candida albicans ChIP-CHIP array |
|
| Samples (6)
|
| GSM338664 |
ChIP-CHIP of Mcm1-TAP in yeast state-2 |
| GSM338665 |
ChIP-CHIP of Mcm1-TAP in hyphal state |
| GSM338856 |
ChIP-CHIP of Mcm1-TAP in hyphal state-2 |
|
| Relations |
| BioProject |
PRJNA110007 |
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