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

doi:10.1534/g3.113.006908

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

School of Medicine

Research output: Contribution to journal › Article › peer-review

39 Scopus citations

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 language	English (US)
Pages (from-to)	1675-1686
Number of pages	12
Journal	G3: Genes, Genomes, Genetics
Volume	3
Issue number	9
DOIs	https://doi.org/10.1534/g3.113.006908
State	Published - 2013

Keywords

Candida glabrata
Expression vector
Inducible
Macrophage
Met3

ASJC Scopus subject areas

Molecular Biology
Genetics
Genetics(clinical)

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10.1534/g3.113.006908

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title = "Expression plasmids for use in Candida glabrata",

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.",

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year = "2013",

doi = "10.1534/g3.113.006908",

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T1 - Expression plasmids for use in Candida glabrata

AU - Zordan, Rebecca E.

AU - Ren, Yuxia

AU - Pan, Shih Jung

AU - Rotondo, Giuseppe

AU - Iluore, Alejandro De Las Peñas

AU - Cormack, Brendan P.

PY - 2013

Y1 - 2013

N2 - 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.

AB - 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.

KW - Candida glabrata

KW - Expression vector

KW - Inducible

KW - Macrophage

KW - Met3

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Expression plasmids for use in Candida glabrata

Abstract

Keywords

ASJC Scopus subject areas

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