Abstract
Inducible promoters are invaluable tools for modulating gene expression (turning transcription on or off) and have been a key approach for ascertaining gene essentiality in Cryptococcus neoformans. Galactose-inducible promoters have been successfully used in Saccharomyces cerevisiae to manipulate heterologous gene expression. Utilizing S. cerevisiae galactose-inducible genes in a BLAST search of the sequenced C. neoformans var. grubii genome, we found three potential galactose-inducible promoters, PGAL1, PGAL7, and PUGE2 that are induced by galactose and repressed by glucose in this variety. This chapter describes how to make a fusion of these promoters with heterologous genes, how to insert the fused gene back into the genome, and how to induce expression during asexual and sexual growth in C. neoformans var. grubii.
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Acknowledgments
This work was supported by an NIH-NIAID grants RO1-AI50184 and RO1-AI072195 to JKL.
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Baker, L.G., Lodge, J.K. (2012). Galactose-Inducible Promoters in Cryptococcus neoformans var. grubii . In: Brand, A., MacCallum, D. (eds) Host-Fungus Interactions. Methods in Molecular Biology, vol 845. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-61779-539-8_14
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DOI: https://doi.org/10.1007/978-1-61779-539-8_14
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