ISOLATION AND CHARACTERIZATION OF THE CANDIDA-ALBICANS PFY1 GENE FOR PROFILIN

We have isolated the Candida albicans gene for profilin, PFY1. Degener ate oligonucleotide primers based on regions of high homology were uti lized to obtain a polymerase chain reaction-amplified copy of the gene . This was then used as a probe to isolate the gene from a C. albicans genomic library. Our studies indicate that the full-length gene is un stable in Escherichia coli. Several clones were sequenced, and the pre dicted amino acid sequence demonstrated homology with profilin protein s from other organisms, most notably Saccharomyces cerevisiae. Norther n analysis revealed that the gene is expressed in C. albicans. Attempt s to express the gene in S. cerevisiae cells were unsuccessful until t he C. albicans promoter was replaced with an S. cerevisiae promoter. F unctional complementation of the gene was demonstrated in S. cerevisia e profilin-requiring cells. Antibodies raised to isolated C. albicans profilin protein recognized a protein of the predicted molecular weigh t when the gene was expressed in S. cerevisiae cells. The sequence of the C. albicans PFY1 gene has been deposited in the Genome Sequence da tabase under Accession Number L3783. (C) 1997 by John Wiley & Sons, Lt d.