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Expression Plasmids for Use in Candida glabrata

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...

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Autores principales: Zordan, Rebecca E., Ren, Yuxia, Pan, Shih-Jung, Rotondo, Giuseppe, Peñas, Alejandro De Las, Iluore, Joseph, Cormack, Brendan P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3789792/
https://www.ncbi.nlm.nih.gov/pubmed/23934995
http://dx.doi.org/10.1534/g3.113.006908
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author Zordan, Rebecca E.
Ren, Yuxia
Pan, Shih-Jung
Rotondo, Giuseppe
Peñas, Alejandro De Las
Iluore, Joseph
Cormack, Brendan P.
author_facet Zordan, Rebecca E.
Ren, Yuxia
Pan, Shih-Jung
Rotondo, Giuseppe
Peñas, Alejandro De Las
Iluore, Joseph
Cormack, Brendan P.
author_sort Zordan, Rebecca E.
collection PubMed
description 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.
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spelling pubmed-37897922013-10-17 Expression Plasmids for Use in Candida glabrata Zordan, Rebecca E. Ren, Yuxia Pan, Shih-Jung Rotondo, Giuseppe Peñas, Alejandro De Las Iluore, Joseph Cormack, Brendan P. G3 (Bethesda) Investigations 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. Genetics Society of America 2013-10-01 /pmc/articles/PMC3789792/ /pubmed/23934995 http://dx.doi.org/10.1534/g3.113.006908 Text en Copyright © 2013 Zordan et al. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigations
Zordan, Rebecca E.
Ren, Yuxia
Pan, Shih-Jung
Rotondo, Giuseppe
Peñas, Alejandro De Las
Iluore, Joseph
Cormack, Brendan P.
Expression Plasmids for Use in Candida glabrata
title Expression Plasmids for Use in Candida glabrata
title_full Expression Plasmids for Use in Candida glabrata
title_fullStr Expression Plasmids for Use in Candida glabrata
title_full_unstemmed Expression Plasmids for Use in Candida glabrata
title_short Expression Plasmids for Use in Candida glabrata
title_sort expression plasmids for use in candida glabrata
topic Investigations
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3789792/
https://www.ncbi.nlm.nih.gov/pubmed/23934995
http://dx.doi.org/10.1534/g3.113.006908
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