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 which differ in their selectable marker - URA3, or the dominant-selectable NAT1 gene, which confers resistance to the drug nourseothricin (Nat). Expression from the twelve resulting plasmids was assessed using GFP as a reporter and flow cytometry or quantitative reverse-transcription PCR (RTqPCR) to assess expression levels. Together this set of plasmids expands the toolkit of expression vectors available for use in C. glabrata.

Expression plasmids for use in Candida glabrata / R.E. Zordan, Y. Ren, S.J. Pan, G. Rotondo, A. De Las Peñas, J. Iluore, B.P. Cormack. - In: G3. - ISSN 2160-1836. - 3:10(2013 Oct 01), pp. 1675-1686.

Expression plasmids for use in Candida glabrata

G. Rotondo;
2013-10-01

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 which differ in their selectable marker - URA3, or the dominant-selectable NAT1 gene, which confers resistance to the drug nourseothricin (Nat). Expression from the twelve resulting plasmids was assessed using GFP as a reporter and flow cytometry or quantitative reverse-transcription PCR (RTqPCR) to assess expression levels. Together this set of plasmids expands the toolkit of expression vectors available for use in C. glabrata.
Candida glabrata; expression vector; macrophage; inducible; MET3
Settore BIO/11 - Biologia Molecolare
Settore BIO/18 - Genetica
Settore BIO/19 - Microbiologia Generale
G3
Article (author)
File in questo prodotto:
File Dimensione Formato  
1675.full.pdf

accesso riservato

Tipologia: Publisher's version/PDF
Dimensione 1.6 MB
Formato Adobe PDF
1.6 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

Caricamento pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/227617
Citazioni
  • ???jsp.display-item.citation.pmc??? 33
  • Scopus 47
  • ???jsp.display-item.citation.isi??? 49
social impact