Study on the relationship between K143Q amino acid substitution in 14α-demethylase and fluconazole resistance to Candida albicans

doi:10.3969/j.issn.1673-8640.2014.07.014

›› 2014, Vol. 29 ›› Issue (7): 750-754.DOI: 10.3969/j.issn.1673-8640.2014.07.014

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Study on the relationship between K143Q amino acid substitution in 14α-demethylase and fluconazole resistance to Candida albicans

LIU Jinyan¹,NI Peihua²,SHI Ce¹,WEI Bing¹,XIANG Mingjie³

1. Department of Radioimmunology and Laboratory Medicine, Luwan Branch, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200020, China;
2.Faculty of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China;
3.Department of Clinical Laboratory, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200020, China

Received:2013-09-22 Online:2014-07-30 Published:2014-07-21

Abstract

Abstract:

Objective　To verify the correlation of K143Q amino acid substitution in 14α-demethylase and fluconzole resistance to Candida albicans. Methods　The recombinant mutant plasmid with A427C in ERG11 gene was firstly constructed by site-directed mutagenesis， and YES2/CT yeast expression plasmid was established by digestion and ligation technique. The heterologous expression in Saccharomyces cerevisiae and drug susceptibility in vitro were performed to analyze the correlation of K143Q amino acid substitution with resistance to fluconazole. Results　The antifungal drug susceptibility test in vitro showed that the constructed plasmid with amino acid substitution transferred in Saccharomyces cerevisiae INVSc1 contributed to increase in fluconazole minimal inhibitory concentration（MIC） for 16 times. Conclusions　The K143Q contributes to increase the resistance to fluconazole in Candida albicans clinical isolates.

Key words: Candida albicans, Fluconazole, ERG11 gene, Missense mutation, Site-directed mutagenesis, Saccharomyces cerevisiae, Heterologous expression

CLC Number:

R446.5

LIU Jinyan,NI Peihua,SHI Ce,WEI Bing,XIANG Mingjie. Study on the relationship between K143Q amino acid substitution in 14α-demethylase and fluconazole resistance to Candida albicans[J]. , 2014, 29(7): 750-754.

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Study on the relationship between K143Q amino acid substitution in 14α-demethylase and fluconazole resistance to Candida albicans

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