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Eukaryotic Cell, June 2006, p. 935-944, Vol. 5, No. 6
1535-9778/06/$08.00+0     doi:10.1128/EC.00028-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of a Copper-Inducible Promoter for Use in Ectopic Expression in the Fungal Pathogen Histoplasma capsulatum{dagger}

Dana Gebhart,{ddagger} Adam K. Bahrami,{ddagger},§ and Anita Sil*

Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California 94143-0414

Received 31 January 2006/ Accepted 10 April 2006

Despite the existence of a number of genetic tools to study the fungal pathogen Histoplasma capsulatum, strategies for conditional gene expression have not been developed. We used microarray analysis to identify genes that are transcriptionally induced or repressed by the addition of copper sulfate (CuSO4) to H. capsulatum yeast cultures. One of these genes, CRP1, encodes a putative copper efflux pump that is significantly induced in the presence of CuSO4. The upstream regulatory region of CRP1 was sufficient to drive copper-regulated expression of two reporter genes, lacZ and the gene encoding green fluorescent protein. Microarray experiments were performed to determine a copper concentration that triggers accumulation of the CRP1 transcript without significant perturbation of global gene expression. These studies show that the CRP1 upstream regulatory region can be used for ectopic expression of heterologous genes in H. capsulatum. Furthermore, they demonstrate the strategic use of microarrays to identify conditional promoters that confer induction in the absence of large-scale shifts in gene expression.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143-0414. Phone: (415) 502-1805. Fax: (415) 476-8201. E-mail: sil{at}cgl.ucsf.edu.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

{ddagger} These two authors contributed equally to this work.

§ Present address: Department of Organismic & Evolutionary Biology, Harvard University, 26 Oxford St., Cambridge, MA 02138.

Eukaryotic Cell, June 2006, p. 935-944, Vol. 5, No. 6
1535-9778/06/$08.00+0     doi:10.1128/EC.00028-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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