CRISPR/Cas9方法失活白念珠菌CaMIT1基因及其突变株表型分析

中国真菌学杂志　 2018, Vol. 13 　Issue (4): 201-207.

CRISPR/Cas9方法失活白念珠菌CaMIT1基因及其突变株表型分析

徐慧慧¹, 蒋伶活^1,2

1. 江南大学生物工程学院粮食发酵工艺与技术国家工程实验室, 无锡 214122;
2. 山东理工大学农业工程与食品科学学院, 淄博 255000

收稿日期:2017-12-20 出版日期:2018-08-28 发布日期:2018-08-28
通讯作者: 蒋伶活,E-mail:linghuojiang@sdut.edu.cn E-mail:linghuojiang@sdut.edu.cn
作者简介:徐慧慧,女(汉族),博士研究生在读.E-mail:xuhuijndx2012@hotmail.com
基金资助:
国家自然科学基金（81371784，81571966）

Inactivation of Candida albicans CaMIT1 gene with CRISPR/Cas9 approach and the phenotype analysis of its homozygous mutant

XU Hui-hui¹, JIANG Ling-huo^1,2

1. School of Biotechnology, the National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122;
2. School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255000

Received:2017-12-20 Online:2018-08-28 Published:2018-08-28

摘要/Abstract

摘要：

目的构建白念珠菌CaMIT1基因突变株，研究CaMIT1基因在白念珠菌中的功能。方法本文采用CRISPR（Clustered Regularly Interspaced Short Palindromic Repeat）/Cas9这种新的方法构建CaMIT1失活突变株，并通过倍比稀释的方法初步研究CaMIT1的功能。结果成功获得了CaMIT1基因的纯合子失活突变株，表型实验结果显示该基因突变株对钙离子、锂离子、十二烷基磺酸钠、克霉唑、酮康唑、Anidualafungin敏感，对刚果红耐受。结论 CaMIT1与白念珠菌的细胞质膜和细胞壁完整性以及钙离子的稳态相关。

关键词: 白念珠菌, CaMIT1, CRISPR, 表型分析, 细胞质膜和细胞壁压力

Abstract:

Objective To construct mutants for C.albicans CaMIT1 and investigate its functions in C. albicans.Methods We constructed the inactivated mutant for CaMIT1 with the novel approach CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat)/Cas9. Serial dilution assay was used to analyze the function of CaMIT1. Results We successfully constructed the homozygous inactivation mutant for CaMIT1, and phenotypic analysis revealed that the homozygous mutant was sensitive to calcium and lithium ions, sodium dodecyl sulfate, clotrimazole, ketoconazole and anidualafungin, but tolerant to Congo Red. Conclusions CaMIT1 is involved in the integrity of plasma membrane and cell wall as well as the calcium homeostasis.

Key words: Candida albicans, CaMIT1, CRISPR, Phenotypic analysis, Plasma membrane and cell wall stress

中图分类号:

R379.4

徐慧慧, 蒋伶活. CRISPR/Cas9方法失活白念珠菌CaMIT1基因及其突变株表型分析[J]. 中国真菌学杂志, 2018, 13(4): 201-207.

XU Hui-hui, JIANG Ling-huo. Inactivation of Candida albicans CaMIT1 gene with CRISPR/Cas9 approach and the phenotype analysis of its homozygous mutant[J]. Chinese Journal of Mycology, 2018, 13(4): 201-207.

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