Expression plasmids for use in Candida glabrata

Rebecca E. Zordan, Yuxia Ren, Shih Jung Pan, Giuseppe Rotondo, Alejandro De Las Peñas Iluore, Brendan P. Cormack

Research output: Contribution to journalArticle

Abstract

We describe a series of CEN/ARS episomal plasmids containing different Candida glabrata promoters, allowing for a range of constitutive or regulated expression of proteins in C. glabrata. The set of promoters includes three constitutive promoters (EGD2pr, HHT2pr, PDC1pr), two macrophage/phagocytosisinduced promoters (ACO2pr, LYS21pr), and one nutritionally regulated promoter (MET3pr). Each promoter was cloned into two plasmid backbones that differ in their selectable marker, URA3, or the dominant-selectable NAT1 gene, which confers resistance to the drug nourseothricin. Expression from the 12 resulting plasmids was assessed using GFP as a reporter and flow cytometry or quantitative reverse-transcription polymerase chain reaction to assess expression levels. Together this set of plasmids expands the toolkit of expression vectors available for use with C. glabrata.

Original languageEnglish (US)
Pages (from-to)1675-1686
Number of pages12
JournalG3: Genes, Genomes, Genetics
Volume3
Issue number9
DOIs
StatePublished - Jan 1 2013

Keywords

  • Candida glabrata
  • Expression vector
  • Inducible
  • Macrophage
  • Met3

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

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  • Cite this

    Zordan, R. E., Ren, Y., Pan, S. J., Rotondo, G., Iluore, A. D. L. P., & Cormack, B. P. (2013). Expression plasmids for use in Candida glabrata. G3: Genes, Genomes, Genetics, 3(9), 1675-1686. https://doi.org/10.1534/g3.113.006908