Disruption of the gene encoding the secreted acid protease (ACP) in the yeast Candida tropicalis.
The gene for the secreted acid protease (ACP), a potential virulence factor of Candida species, was inactivated in Candida tropicalis by gene disruption. The disruption was performed by cotransformation of an ade2 C. tropicalis mutant with a linear DNA fragment carrying a deletion in ACP, and the replicative vector pMK16 which carries a selectable ADE2 gene marker. Few of the transformants exhibited lower protease secretion levels and were shown to have one deleted and one unaffected ACP copy, since C. tropicalis is a diploid yeast. These transformants were rendered homozygotic for this deletion by mild UV-treatment. One of the homozygotic acp deletion mutants obtained was completely devoid of extracellular protease activity and grew poorly on bovine serum albumin-containing medium. This mutant could be complemented by an ACP fragment inserted in pMK16, but also by an acid protease gene isolated from C. parapsilosis.
Sanglard
D
D
Institut für Biotechnologie, ETH-Hönggerberg, Zürich, Switzerland.
Togni
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G
de Viragh
P A
PA
Monod
M
M
D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000378:metabolism; D002175:Candida / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D002872:Chromosome Deletion; D003470:Culture Media; D015870:Gene Expression / Q000235:genetics; D009154:Mutation; D012710:Serum Albumin, Bovine
D003470:Culture Media; D012710:Serum Albumin, Bovine; D016282:Aspartic Acid Endopeptidases