Analysis of gene expression in filamentous cells of Candida albicans grown on agar plates
Main Article Content
Keywords
Candida albicans, filamentation, RNA extraction, qRT-PCR, RNAseq
Abstract
Candida albicans (C. albicans) is a commensal organism of the human gastrointestinal and genitourinary tracts. C. albicans is also a major human pathogen, causing disease ranging from cutaneous infections to lethal systemic disease. The ability of this fungus to switch between yeast and filamentous forms of growth has long been linked to its pathogenesis. Filamentation can be induced by a variety of distinct environmental cues and can occur in either liquid or solid media. While some evidence suggests that there are differences between filamentation in solid and liquid media, gene expression analysis of filamentation in C. albicans has focused strictly on cells grown in liquid media. We have developed a method for analyzing gene expression of filamentous cells grown on solid induction media at early stages of filamentation, establishing cell plating densities, ideal collection times, and collection techniques. We have also demonstrated the utility of the approach not only in qRT-PCR assays, but high-throughput RNAseq assays as well. These assays will allow for comparison studies of C. albicans filamentation initiation in solid and liquid media.
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References
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3. Zheng X, Wang Y, Wang Y. Hgc1, a novel hypha-specific G1 cyclin-related protein regulates Candida albicans hyphal morphogenesis. EMBO J. 2004;23(8):1845-56. doi: 10.1038/sj.emboj.7600195. PubMed PMID: 15071502; PubMed Central PMCID: PMCPMC394249.
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6. Lee KL, Buckley HR, Campbell CC. An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida albicans. Sabouraudia. 1975;13(2):148-53. PubMed PMID: 808868.
7. Chen H, Fujita M, Feng Q, Clardy J, Fink GR. Tyrosol is a quorum-sensing molecule in Candida albicans. Proc Natl Acad Sci U S A. 2004;101(14):5048-52. doi: 10.1073/pnas.0401416101. PubMed PMID: 15051880; PubMed Central PMCID: PMCPMC387371.
8. Hornby JM, Jensen EC, Lisec AD, Tasto JJ, Jahnke B, Shoemaker R, et al. Quorum sensing in the dimorphic fungus Candida albicans is mediated by farnesol. Appl Environ Microbiol. 2001;67(7):2982-92. doi: 10.1128/AEM.67.7.2982-2992.2001. PubMed PMID: 11425711; PubMed Central PMCID: PMCPMC92970.
9. Staab JF, Bradway SD, Fidel PL, Sundstrom P. Adhesive and mammalian transglutaminase substrate properties of Candida albicans Hwp1. Science. 1999;283(5407):1535-8. PubMed PMID: 10066176.
10. Braun BR, Head WS, Wang MX, Johnson AD. Identification and characterization of TUP1-regulated genes in Candida albicans. Genetics. 2000;156(1):31-44. PubMed PMID: 10978273; PubMed Central PMCID: PMCPMC1461230.
11. Birse CE, Irwin MY, Fonzi WA, Sypherd PS. Cloning and characterization of ECE1, a gene expressed in association with cell elongation of the dimorphic pathogen Candida albicans. Infect Immun. 1993;61(9):3648-55. PubMed PMID: 8359888; PubMed Central PMCID: PMCPMC281060.
12. Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, et al. The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol. 2006;7:3. Epub 2006/02/02. doi: 10.1186/1471-2199-7-3. PubMed PMID: 16448564; PubMed Central PMCID: PMCPMC1413964.
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Published
Jan 23, 2018
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Creger PE, Blankenship JR. Analysis of gene expression in filamentous cells of Candida albicans grown on agar plates. J Biol Methods [Internet]. 2018 Jan. 23 [cited 2024 Apr. 19];5(1):e84. Available from: https://jbmethods.org/index.php/jbm/article/view/211
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Author Biography
Jill R. Blankenship, University of Nebraska at Omaha
Biology Department, Assistant professor
