Article (Scientific journals)
New disruption cassettes for rapid gene disruption and marker rescue in the yeast Yarrowia lipolytica
Fickers, Patrick; Le Dall, M. T.; Gaillardin, C. et al.
2003In Journal of Microbiological Methods, 55 (3), p. 727-737
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Keywords :
gene disruption; Cre-lox; recombinase; Yarrowia lipolytica
Abstract :
[en] Yarrowia lipolytica is one of the most extensively studied nonconventional yeasts. Unfortunately, few methods for gene disruption have been reported for this yeast, and all of them are time-consuming and laborious. The functional analysis of unknown genes requires powerful disruption methods. Here, we describe such a new method for rapid gene disruption in Y lipolytica. This knockout system combines SEP method and the Cre-lox recombination system, facilitating efficient marker rescue. Versatility was increased by using both auxotrophic markers like ylURA3 and ylLEU2, as well as the antibiotic resistance marker hph. The hph marker, which confers resistance to hygromycin-B, allows gene disruption in a strain lacking any conventional auxothrophic marker. The disruption cassette was shown to integrate at the correct locus at an average frequency of 45%. Upon expression of Cre recombinase, the marker was excised at a frequency of 98%, by recombination between the two lox sites. This new method for gene disruption is an ideal tool for the functional analysis of gene families, or for creating large-scale mutant collections in general. (C) 2003 Elsevier B.V. All rights reserved.
Disciplines :
Microbiology
Biochemistry, biophysics & molecular biology
Author, co-author :
Fickers, Patrick ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Le Dall, M. T.
Gaillardin, C.
Thonart, Philippe ;  Université de Liège - ULiège > Gembloux Agro-Bio Tech > Gembloux Agro-Bio Tech - Biochimie et microbiologie industrielles
Nicaud, J. M.
Language :
English
Title :
New disruption cassettes for rapid gene disruption and marker rescue in the yeast Yarrowia lipolytica
Publication date :
December 2003
Journal title :
Journal of Microbiological Methods
ISSN :
0167-7012
eISSN :
1872-8359
Publisher :
Elsevier Science Bv, Amsterdam, Netherlands
Volume :
55
Issue :
3
Pages :
727-737
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
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