title: Application of the Cre-loxP system for multiple gene disruption in the yeast Kluyveromyces marxianus.
abstract: 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 beta-galactosidase activity.
authors.affiliation: Institute of Biotechnology and Bioengineering (IBB), Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, Braga, Portugal.
authors.forename: Orquídea
authors.initials: O
authors.lastname: Ribeiro
authors.forename: Andreas K
authors.initials: AK
authors.lastname: Gombert
authors.forename: José A
authors.initials: JA
authors.lastname: Teixeira
authors.forename: Lucília
authors.initials: L
authors.lastname: Domingues
mesh_terms: D001287:Attachment Sites, Microbiological / Q000235:genetics*; D015415:Biomarkers / Q000378:metabolism; D002244:Carbon / Q000494:pharmacology; D018274:Electroporation; D018390:Gene Targeting / Q000379:methods*; D005800:Genes, Fungal / Q000235:genetics*; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D007716:Kluyveromyces / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D015415:Biomarkers; D002244:Carbon; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase