title: Development of Streptococcus thermophilus lacZ as a reporter gene for Candida albicans.
abstract: The study of gene regulation in many organisms has been facilitated by the development of reporter genes. The authors report the use of lacZ from Streptococcus thermophilus, a gene encoding a beta-galactosidase, as a reporter for the fungal pathogen Candida albicans. As test cases, Strep. thermophilus lacZ was placed under control of three different C. albicans promoters: MAL2 (maltase), inducible by maltose; HWP1 (hyphal cell wall protein), induced by conditions that promote filamentous growth; and ACT1 (actin). These constructs were each integrated into the C. albicans genome and beta-galactosidase activity was readily detected from these strains, but only under the appropriate growth conditions. Beta-galactosidase activity could be detected by several methods: quantitative liquid assays using permeabilized cells, colorimetric assays of colonies replicated to paper filters, and in situ coloration of colonies growing on medium containing the indicator X-Gal. These results show the usefulness of STREP: thermophilus lacZ as a monitor of gene regulation in this medically important yeast.
authors.affiliation: Department of Microbiology and Immunology, University of California, San Francisco, CA 94143-0414, USA1.
authors.forename: M Andrew
authors.initials: MA
authors.lastname: Uhl
authors.affiliation: Department of Microbiology and Immunology, University of California, San Francisco, CA 94143-0414, USA1.
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Johnson
mesh_terms: D000199:Actins / Q000235:genetics; D002176:Candida albicans / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins; D017930:Genes, Reporter; D007763:Lac Operon; D008562:Membrane Glycoproteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013291:Streptococcus / Q000235:genetics*; D000520:alpha-Glucosidases / Q000235:genetics; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C404682:ACT1 protein, Candida albicans; D000199:Actins; D005656:Fungal Proteins; C118102:HWP1 protein, Candida albicans; D008562:Membrane Glycoproteins; D000520:alpha-Glucosidases; D001616:beta-Galactosidase