| 白念珠菌14-α脱甲基酶K143Q氨基酸置换与氟康唑耐药形成的相关性研究 |
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| 引用本文： | 刘锦燕,;倪培华,;史册,;魏冰,;项明洁.白念珠菌14-α脱甲基酶K143Q氨基酸置换与氟康唑耐药形成的相关性研究[J].检验医学,2014(7):750-754. |
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| 作者姓名： | 刘锦燕 ;倪培华 ;史册 ;魏冰 ;项明洁 |
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| 作者单位： | [1]上海交通大学医学院附属瑞金医院卢湾分院放免检验科,上海200020; [2]上海交通大学医学院附属瑞金医院检验系,上海200025; [3]上海交通大学医学院附属瑞金医院检验科,上海200020 |
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| 基金项目： | 上海市科委课题（114119b0500）; 上海交通大学医学院课题（09XJ21036）; 上海市卫生局课题（2009239） |
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| 摘 要： | 目的研究临床耐氟康唑白念珠菌14-α脱甲基酶的K143Q氨基酸置换与氟康唑耐药形成的关系。方法运用体外定点突变技术构建ERG11基因A427C突变型重组质粒,并利用酶切、连接构建YES2/CT酵母表达质粒。通过构建成的表达质粒在酿酒酵母中的异源性表达及体外药物敏感性表型的检测,分析其致K143Q氨基酸置换与白念珠菌对氟康唑产生耐药的关系。结果真菌体外药物敏感性表型检测显示,含氨基酸置换的表达质粒转染于酿酒酵母INVSc1后,氟康唑最低抑菌浓度(MIC)增加16倍。结论 K143Q可增强白念珠菌对氟康唑的耐药性。
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| 关 键 词： | 白念珠菌 氟康唑 ERG11基因 错义突变 定点突变技术 酿酒酵母 异源性表达 |
Study on the relationship between K143Q amino acid substitution in 14α-demethylase and fluconazole resistance to Candida albicans |
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| Institution: | LIU Jinyan, NI Peihua, SHI Ce, WEI Bing, XIANG Mingjie( 1. Department of Radioimmunology and Laboratory Medicine, Lawan 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) |
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| Abstract: | Objective To verify the correlation of K143 Q amino acid substitution in 14α-demethylase and fluconzole resistance to Candida albicans. Methods The recombinant mutant plasmid with A427 C 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 K143 Q 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 K143 Q contributes to increase the resistance to fluconazole in Candida albicans clinical isolates. |
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| Keywords: | Candida albicans Fluconazole ERG11 gene Missense mutation Site-directed mutagenesis Saccharomyces cerevisiae Heterologous expression |
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