白念珠菌新型耐药基因MXR1的敲除与鉴定

中国真菌学杂志　 2013, Vol. 8 　Issue (2): 69-73.

白念珠菌新型耐药基因MXR1的敲除与鉴定

谭宏月¹, 李平¹, 胡婵¹, 朱红梅¹, 顾军², 温海¹

1. 上海长征医院皮肤病与真菌病研究所全军真菌病重点实验室第二军医大学附属长征医院皮肤科, 上海 200003;
2. 第二军医大学附属长海医院皮肤科, 上海 200433

收稿日期:2012-08-22 出版日期:2013-04-28 发布日期:2013-04-28
通讯作者: 顾军,E-mail:gujun79@163.com;温海,E-mail:wenhai98@sohu.com E-mail:gujun79@163.com;wenhai98@sohu.com
作者简介:谭宏月,女 (汉族),博士研究生在读,住院医师.E-mail:sabrina_tan@126.com
基金资助:
国家自然科学基金(30872276)

Construction of knockout strains of MXR1 gene in Candida albicans

TAN Hong-yue¹, LI Ping¹, HU Chan¹, ZHU Hong-mei¹, GU Jun², WEN Hai¹

1. Institute of dermatology and fungal disease, Key Laboratory of the army fungal disease, Department of dermatology, Changzheng Hospital, Shanghai 200003;
2. Department of Dermatology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China

Received:2012-08-22 Online:2013-04-28 Published:2013-04-28

摘要/Abstract

摘要： 目的构建用于白念珠菌MXR1基因敲除的载体质粒,并通过Ura-Blaster策略敲除MXR1两条等位基因。方法分别扩增白念珠菌MXR1基因ORF两侧上下游的片段,通过酶切与连接反应,将上下游片段分别插入到p5921质粒的hisG-URA3-hisG盒两端,从而形成MXR1敲除载体质粒pUC-MXR1-URA3。通过Ura-Blaster策略将载体质粒转染到白念珠菌RM1000内,并采用PCR和Southern-blot杂交方法鉴定各步转染、复筛所得的阳性克隆。结果成功获得MXR1基因缺失的菌株。结论 MXR1基因缺失菌株的构建,有助于深入研究白念珠菌耐药机制。

关键词: 白念珠菌, MXR1基因, 基因敲除

Abstract: Objective To construct the plasmid for MXR1 disruption, and knockout the MXR1 gene in Candida albicans.Methods Up and down streams of MXR1 gene ORF were amplified and inserted into the p5921 plasmid.MXR1 gene was disrupted according to the Ura-Blaster method. The constructed strains were identified by PCR and Southern-blot methods.Results The knockout plasmid pUC-MXR1-URA3 and MXR1 disruption strains were successfully constructed.Conclusions The MXR1 disruption strains are helpful for the study of the drug-resistance mechanism.

Key words: Candida albicans, MXR1 gene, gene disruption

中图分类号:

R379.4

谭宏月, 李平, 胡婵, 朱红梅, 顾军, 温海. 白念珠菌新型耐药基因MXR1的敲除与鉴定[J]. 中国真菌学杂志, 2013, 8(2): 69-73.

TAN Hong-yue, LI Ping, HU Chan, ZHU Hong-mei, GU Jun, WEN Hai. Construction of knockout strains of MXR1 gene in Candida albicans[J]. Chinese Journal of Mycology, 2013, 8(2): 69-73.

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