Mutants of Candida albicans hypersensitive to calcofluor white display susceptibility to antifungal drugs.
An increased infection incidence of Candida albicans (most common human fungal pathogen) contributes to the need of further functional genetic studies and development of new antifungal drugs. We developed a method to create mutants of C. albicans using an antisense cDNA library to interfere with gene expression, followed by screening for hypersensitivity to Calcofluor White (CFW) and the antifungal drugs caspofungin and itraconazole. Mutants with these properties have with a high probability defects in cell-wall integrity. Fifty out of 200 transformant colonies analyzed (25%) showed hypersensitivity to CFW compared with the parental strain C. albicans CAI-4. Most of those CFW-hypersensitive mutants further displayed the susceptibility to antifungal drugs itraconazole and caspofungin using microbroth dilution method M27-A and an agar-diffusion test. The mutants obtained through this procedure could provide a potential model for screening antifungal pro-drugs which show weak action when standard C. albicans strain is used and may also aid in further identifying genes involved in cell integrity. In addition, we describe the effect of varying several parameters in electroporation transformation, including treatment with lithium acetate, upon the efficiency of transformation in C. albicans.
He
Zhu-Mei
ZM
Key Laboratory of Gene Engineering of the Ministry of Education/Guangdong Provincial Research Center for Severe Infectious Disease Prevention and Control Technology, Sun Yat-Sen University, Guangzhou, 510275, P.R. China. lsshezm@mail.sysu.edu.cn
Chen
Jing
J
Li
Hong-Zhou
HZ
Luo
Di-Qing
DQ
Yang
Wei
W
D000935:Antifungal Agents / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects* / Q000235:genetics*; D000077336:Caspofungin; D002473:Cell Wall / Q000187:drug effects / Q000235:genetics; D054714:Echinocandins / Q000494:pharmacology*; D055666:Lipopeptides; D008826:Microbial Sensitivity Tests; D009154:Mutation
D000935:Antifungal Agents; D054714:Echinocandins; D055666:Lipopeptides; D000077336:Caspofungin