ldentification of differentially expressed genes in Candida albicans during dimorphic transition using differential display reverse transcription polymerase chain reaction DDRT-PCR

Seow Heng Fong, . and Lim Crystale Siew Ying, . and Chong Pei Pei, . and Wong Won Fen, . and Yonq Phelim Voon Chen, . (2005) ldentification of differentially expressed genes in Candida albicans during dimorphic transition using differential display reverse transcription polymerase chain reaction DDRT-PCR. [Proceedings Paper]

Superficial and invasive candidiasis had increased remarkably over the past few years. Candida albicans is the most common type of Candida species. lt possesses a variety of virulence attributes such as adhesion to host cell ability to secrete hydrolytic enzymes but the most significant virulent factor is the ability to form hyphae. The molecular basis of C.albicans morphology switching is still an unsolved mystery despite many previous research efforts. Thus this study was aimed at identifying the gene expression of Candida albicans during its conversion from non-pathogenic yeast to pathogenic hyphal form by utilizing a differential display reverse transcribed polymerase chain reaction DDRT-PCR method. mRNA was extracted from both yeast and hyphal cells at different time point of hyphal elongation. RNA image kit1 was used to carry out DDRT-PCR experiments. The differentially expressed bands were determined from the autoradiogram recovered from the gel and sequenced. The results were confirmed using Northern hybridization method and semi-quantitative RT-PCR. Five genes involved in the cAMP signaling were found to be upregulated in the hyphal cells. Genes involved in cell cycle regulation genotypic switching starvation and glucose sensing were also expressed during dimorphic transition. lnterestingly genes encoding in mitochondria transporter riboflavin synthase glutamate synthase and methionine permease were discovered to be differentially expressed. ln addition three hyphothetical genes with unknown function were also found. Further studies on the potential direct functional roles that these newly discovered genes play in dimorphic transition would lead to a better understanding of hyphae formation mechanism and pathogenesis of Candida albicans.

Full text available from: Official URL: https://docs.google.com/file/d/0B7urwQoldOjeaW5kTm...

Abstract

Superficial and invasive candidiasis had increased remarkably over the past few years. Candida albicans is the most common type of Candida species. lt possesses a variety of virulence attributes such as adhesion to host cell ability to secrete hydrolytic enzymes but the most significant virulent factor is the ability to form hyphae. The molecular basis of C.albicans morphology switching is still an unsolved mystery despite many previous research efforts. Thus this study was aimed at identifying the gene expression of Candida albicans during its conversion from non-pathogenic yeast to pathogenic hyphal form by utilizing a differential display reverse transcribed polymerase chain reaction DDRT-PCR method. mRNA was extracted from both yeast and hyphal cells at different time point of hyphal elongation. RNA image kit1 was used to carry out DDRT-PCR experiments. The differentially expressed bands were determined from the autoradiogram recovered from the gel and sequenced. The results were confirmed using Northern hybridization method and semi-quantitative RT-PCR. Five genes involved in the cAMP signaling were found to be upregulated in the hyphal cells. Genes involved in cell cycle regulation genotypic switching starvation and glucose sensing were also expressed during dimorphic transition. lnterestingly genes encoding in mitochondria transporter riboflavin synthase glutamate synthase and methionine permease were discovered to be differentially expressed. ln addition three hyphothetical genes with unknown function were also found. Further studies on the potential direct functional roles that these newly discovered genes play in dimorphic transition would lead to a better understanding of hyphae formation mechanism and pathogenesis of Candida albicans.

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Item Type:	Proceedings Paper
AGROVOC Term:	Candida albicans
AGROVOC Term:	Identification
AGROVOC Term:	PCR
AGROVOC Term:	Polymerase chain reaction
AGROVOC Term:	Yeasts
AGROVOC Term:	Hyphae
AGROVOC Term:	Cloning
AGROVOC Term:	Gene expression
AGROVOC Term:	Genes
Geographical Term:	MALAYSIA
Depositing User:	Ms. Suzila Mohamad Kasim
Last Modified:	24 Apr 2025 05:16
URI:	http://webagris.upm.edu.my/id/eprint/13815

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