Galactose-inducible promoters in Cryptococcus neoformans var. grubii

Lorina G. Baker, Jennifer K. Lodge

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

3 Scopus citations

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, P GAL1, P GAL7, and P UGE2 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.

Original languageEnglish
Title of host publicationHost-Fungus Interactions
Subtitle of host publicationMethods and Protocols
EditorsAlexandra Brand, Donna MacCallum
Pages211-226
Number of pages16
DOIs
StatePublished - Mar 16 2012

Publication series

NameMethods in Molecular Biology
Volume845
ISSN (Print)1064-3745

Keywords

  • Galactose induction
  • Glucose repression
  • Mating
  • Native galactose promoters
  • qPCR

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