Characterization of the Candida albicans TRP1 gene and construction of a homozygous trp1 mutant by sequential co-transformation.
The Candida albicans TRP1 gene has been isolated by complementation of an Escherichia coli trpC mutant. Sequence analysis has revealed a single ORF (open reading frame) of 678 nucleotides (nt). The amino acid (aa) sequence deduced from this coding region demonstrates a high degree of homology with PRAI (phosphoribosylanthranilate isomerase) enzymes of other fungi, as well as bacterial species. The gene is also analogous to other yeast TRP1 genes in that it encodes a unifunctional enzyme, whereas TRP1 in filamentous fungi encodes a tri-functional enzyme. Both chromosomal copies of the gene were disrupted by sequential integrative transformation employing co-transformation of an ade1 mutant in order to create a homozygous auxotrophic trp1,ade1 C. albicans strain. This double auxotroph was used to test the ability of the Saccharomyces cerevisiae TRP1 gene to complement the C. albicans trp1 mutation; no expression of the S. cerevisiae gene was detectable.
Department of Microbial Molecular Biology, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ 08543-4000.
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Ostrander
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Gorman
D019747:Aldose-Ketose Isomerases; D000595:Amino Acid Sequence; D001483:Base Sequence; D002176:Candida albicans / Q000235:genetics*; D003001:Cloning, Molecular; D004271:DNA, Fungal; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D006720:Homozygote; D008969:Molecular Sequence Data; D009154:Mutation; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D014170:Transformation, Genetic
D004271:DNA, Fungal; D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; D019747:Aldose-Ketose Isomerases; C490353:TRP1 protein, S cerevisiae