[Cloning and expression of the mutant 14alpha-demethylase containing the alteration of H310D in Candida albicans and the preliminary study on its functions].
The method of recombinant PCR was employed to obtain H310D mutation in 14alpha-demethylase in Candida albicans). The expression vector pYCYPS1M harboning the mutant CYP1M gene was constructed. The mutant CYPS1 was expressed in Sacchromyces cerevisiae induced by 2% galactose. At last the MICs of yeasts to Fluconazole were determined by broth microdilution testing. Expressed protein was 15% of the total microsome proteins. The MICs of yeasts expressed the mutant CYP51 were 4-fold as that of the yeasts expressed the wild CYP51. The increase of the MICs was due to the alteration H310D in CYP51. It could be concluded that the residue of H310 was important to the function of CYPS1.
Gao
Pinghui
P
Department of Pharmacology, Second Military Medical University, Shanghai 200433, China. gaoph@yahoo.com
Cao
Yongbing
Y
Xu
Zheng
Z
Liu
Hongtao
H
Jiang
Yuanying
Y
D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D020125:Mutation, Missense; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
D005656:Fungal Proteins; D003577:Cytochrome P-450 Enzyme System