Mutants of <Emphasis Type="Italic">Candida albicans</Emphasis> hypersensitive to calcofluor white display susceptibility to antifungal drugs |
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Authors: | Zhu-Mei He Jing Chen Hong-Zhou Li Di-Qing Luo Wei Yang |
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Institution: | 1.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,P.R. China;2.First Affiliated Hospital,Sun Yat-Sen University,Guangzhou,P.R. China |
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Abstract: | 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. |
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