Objective To construct deletion mutants for Candida albicans CaCSC1 and explore its functions.Methods The CaCSC1 knockout cassette containing the SAT1 flipper was PCR amplified using a pair of long primers.The cassette DNA was transformed into the wild type CAI4 strain,and genotypes of correct mutant strains were confirmed by PCR.Phenotypes of gene deletion mutants were screened by serial dilution assay.Results Gene deletion mutants of CaCSC1 were successfully constructed.Deletion mutants of (aCSC1 were sensitive to ketoconazole and tolerant against Zn2+,Mn2+ and Calcofluor white.Conclusions CaCsc1 participates in the regulation of zinc and magnesium homeostasis as well as plasma membrane and cell wall integrity in Candida albicans.%目的 构建白念珠菌CaCSC1基因的基因缺失株,并初步探究它的功能.方法 设计长引物,通过PCR直接扩增出带有SAT1 flipper的CaCSC1基因敲除盒,转化到野生型白念珠菌菌株CAI4,并通过PCR鉴定出基因型正确的转化子,得到CaCSC1的基因缺失株.最后,通过倍比稀释的方法进行表型筛选.结果 成功得到CaCSC1基因的缺失菌株.基因缺失菌株对酮康唑(Ketoconazole)敏感,对Zn2+、Mn2+和荧光增白剂(Calcofluor white)表现出耐受性.结论 CaCsc1蛋白参与对锌离子和锰离子的稳态调控,和细胞膜和细胞壁的完整性也相关.
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