Development of Streptococcus thermophilus lacZ as a reporter gene for Candida albicans.
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.
Uhl
M Andrew
MA
Department of Microbiology and Immunology, University of California, San Francisco, CA 94143-0414, USA1.
Johnson
Alexander D
AD
Department of Microbiology and Immunology, University of California, San Francisco, CA 94143-0414, USA1.
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
C404682:ACT1 protein, Candida albicans; D000199:Actins; D005656:Fungal Proteins; C118102:HWP1 protein, Candida albicans; D008562:Membrane Glycoproteins; D000520:alpha-Glucosidases; D001616:beta-Galactosidase