Application of the Cre- loxP system for multiple gene disruption in the yeast Kluyveromyces marxianus

• Published in: Journal of biotechnology 2007-08, Vol.131 (1), p.20-26
• Main Authors: Ribeiro, Orquídea, Gombert, Andreas K., Teixeira, José A., Domingues, Lucília
• Format: Article
• Language: English
• Subjects: Attachment Sites, Microbiological - genetics; bacteriophages; beta-galactosidase; beta-Galactosidase - metabolism; Biological and medical sciences; Biomarkers - metabolism; Biotechnology; Carbon - pharmacology; Cre- loxP system; Electroporation; Fundamental and applied biological sciences. Psychology; Gene disruption; gene targeting; Gene Targeting - methods; Gene targeting efficiency; genes; Genes, Fungal - genetics; genetic recombination; Integrases - genetics; Integrases - metabolism; Kluyveromyces - drug effects; Kluyveromyces - genetics; Kluyveromyces - growth & development; Kluyveromyces lactis; Kluyveromyces marxianus; multiple gene disruption; mutants; mutation; Mutation - genetics; Polymerase Chain Reaction; β-Galactosidase
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2007.05.027

The yeast Kluyveromyces marxianus presents several interesting features that make this species a promising industrial yeast for the production of several compounds. In order to take full advantage of this yeast and its particular properties, proper tools for gene disruption and metabolic engineering are needed. The Cre- loxP system is a very versatile tool that allows for gene marker rescue, resulting in mutant strains free of exogenous selective markers, which is a very important aspect for industrial application. As the Cre- loxP system works in some non-conventional yeasts, namely Kluyveromyces lactis, we wished to know whether it also works in K. marxianus. Here, we report the validation of this system in K. marxianus CBS 6556, by disrupting two copies of the LAC4 gene, which encodes a β-galactosidase activity.