title: Gene Expression and Identification Related to Fluconazole Resistance of Candida glabrata Strains.
abstract: OBJECTIVE
  Candida glabrata has become one of the most common causes of Candida bloodstream infections worldwide. Some strains of C. glabrata may be intermediately resistant to all azoles. The several possible mechanisms of azole resistance have been reported previously, but the exact resistant mechanism is not clear. In this study, we identified differentially expressed genes (DEGs) of fluconazole-resistant C. glabrata and compared the gene expression of fluconazole-resistant strains with that of fluconazole-susceptible strains to identify gene corresponding to fluconazole resistance.
  
  
  METHODS
  Using antifungal susceptibility test, several C. glabrata strains were selected and used for further study. The expression of CgCDR1 and CgCDR2 genes was investigated by slot hybridization against fluconazole-susceptible, -resistant, and resistant-induced strains. In addition, ERG3 and ERG11 genes were sequenced to analyze DNA base substitution. DEGs were identified by reverse transcription-polymerase chain reaction using DEG kit composed of 120 random primers.
  
  
  RESULTS
  In slot hybridization, CgCDR1 gene was expressed more than CgCDR2 gene in resistant strains. Though base substitution of ERG11 and ERG3 genes was observed in several base sequences, just one amino acid change was identified in resistant strain. In the results of reverse transcription-polymerase chain reaction, 44 genes were upregulated and 34 genes were downregulated. Among them, adenosine triphosphate-binding cassette transporter-related genes, fatty acid desaturase, lyase, and hypothetical protein genes were upregulated and aldehyde dehydrogenase, oxidoreductase, and prohibitin-like protein genes were downregulated. Other DEGs were also identified.
  
  
  CONCLUSIONS
  This study showed that CgCDR1 gene was more closely related to fluconazole resistance of C. glabrata than CgCDR2 gene. In addition, several other genes related with fluconazole resistance of C. glabrata were identified.
authors.affiliation: Division of Antimicrobial Resistance, Center for Infectious Diseases, National Institute of Health, Korea Centers for Disease Control & Prevention, Seoul, Korea.
authors.forename: Jae Il
authors.initials: JI
authors.lastname: Yoo
authors.forename: Chi Won
authors.initials: CW
authors.lastname: Choi
authors.forename: Kyeong Min
authors.initials: KM
authors.lastname: Lee
authors.forename: Yeong Seon
authors.initials: YS
authors.lastname: Lee
keywords: Candida glabrata; CgCDR1; CgCDR2; fluconazole resistance; gene identification