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research
Cassettes for PCR-mediated gene tagging in Candida albicans utilizing nourseothricin resistance
Authors
Bain
Basso
+36 more
Bates
Bates
Beckerman
Berman
Brand
Calderone
Cannon
Cheng
Fonzi
Gerami-Nejad
Gerami-Nejad
Gietz
Gillum
Hernday
Lavoie
Linder
Murad
Negredo
Niimi
Noble
Noble
Odds
Perlroth
Reijnst
Reuss
Roemer
Schaub
Shen
Staab
Staab
Wach
Warenda
Wilson
Wilson
Wirsching
Wirsching
Publication date
22 November 2013
Publisher
'Wiley'
Doi
Cite
Abstract
addresses: Biosciences, College of Life and Environmental Sciences, University of Exeter, UK.types: Journal Article; Research Support, Non-U.S. Gov't; Validation StudiesThis is the author's post-print version of an article published in Yeast, 2011, Vol. 28, Issue 12, pp. 833 – 841 Copyright © Wiley-Blackwell 2011. The definitive version is available at www3.interscience.wiley.comIn recent years a number of molecular tools have been reported for use in the human fungal pathogen Candida albicans, including PCR-mediated approaches for gene disruption, conditional expression and epitope tagging. Traditionally these methods have utilized auxotrophic markers; however, the availability of auxotrophic markers can be limiting and in some instances their use may also impact on the interpretation of results. As a result, the use of positive selection markers has now become more commonplace. Here we report the development and validation of a set of cassettes for PCR-mediated gene tagging and overexpression studies utilizing the nourseothricin resistance (CaNAT1) positive selection marker. In particular we have produced cassettes containing yeast-enhanced GFP, YFP, CFP, RFP and a combined V5-6xHis epitope tag. The cassettes are engineered for use in PCR-mediated gene tagging strategies where insertion is targeted to the 3' end of the gene of interest. In addition, to facilitate protein functional analysis and genetic suppression studies through the use of overexpression, we have also constructed a promoter replacement cassette containing the ENO1 promoter which is known to be expressed at a high level. These cassettes expand on the range of molecular tools available for working with C. albicans and may also be used in other Candida species that display sensitivity to nourseothricin
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info:doi/10.1002%2Fyea.1910
Last time updated on 22/03/2019
Supporting member
Open Research Exeter
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oai:ore.exeter.ac.uk:10871/139...
Last time updated on 15/12/2013