Expression plasmids for use in Candida glabrata

G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908.

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/phagocytosis-induced 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.

Keywords: Candida glabrata; MET3; expression vector; inducible; macrophage.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Candida glabrata / genetics*
  • Candida glabrata / metabolism
  • DNA, Recombinant / genetics*
  • DNA, Recombinant / metabolism
  • Genes, Reporter
  • Genetic Markers
  • Genetic Vectors / genetics*
  • Genetic Vectors / metabolism
  • Plasmids / genetics*
  • Plasmids / metabolism
  • Promoter Regions, Genetic

Substances

  • DNA, Recombinant
  • Genetic Markers