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Genetics and Molecular Biology |
Department of Microbiology and Immunology, University of California, San Francisco, CA 94143-0414, USA1
Author for correspondence: M. Andrew Uhl. Tel:+1 415 502 0859. Fax: +1 415 476 8201. e-mail: uhl{at}socrates.ucsf.edu
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 ß-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 ß-galactosidase activity was readily detected from these strains, but only under the appropriate growth conditions. ß-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.
Keywords: ß-galactosidase, yeast, fungal pathogen, gene regulation
a Present address: Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA.
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