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Isolation and Characterization of the Candida albicans PFY1 Gene for Profilin
Author(s) -
Ostrander Darin B.,
Gorman Jessica A.
Publication year - 1997
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(199707)13:9<871::aid-yea127>3.0.co;2-2
Subject(s) - biology , gene , profilin , candida albicans , homology (biology) , microbiology and biotechnology , saccharomyces cerevisiae , complementation , genetics , corpus albicans , nucleic acid sequence , mutant , actin cytoskeleton , cytoskeleton , cell
We have isolated the Candida albicans gene for profilin, PFY1 . Degenerate oligonucleotide primers based on regions of high homology were utilized 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 unstable in Escherichia coli . Several clones were sequenced, and the predicted amino acid sequence demonstrated homology with profilin proteins from other organisms, most notably Saccharomyces cerevisiae . Northern analysis revealed that the gene is expressed in C. albicans . Attempts to express the gene in S. cerevisiae cells were unsuccessful until the C. albicans promoter was replaced with an S. cerevisiae promoter. Functional complementation of the gene was demonstrated in S. cerevisiae profilin‐requiring cells. Antibodies raised to isolated C. albicans profilin protein recognized a protein of the predicted molecular weight when the gene was expressed in S. cerevisiae cells. The sequence of the C. albicans PFY1 gene has been deposited in the Genome Sequence database under Accession Number L3783. © 1997 John Wiley & Sons, Ltd.