1. Selectable marker recycling in the nonconventional yeast Xanthophyllomyces dendrorhous by transient expression of Cre on a genetically unstable vector.
- Author
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Zhang N, Li J, Li F, and Wang S
- Subjects
- Gene Knockout Techniques, Integrases genetics, Recombination, Genetic, Selection, Genetic, Yarrowia genetics, Basidiomycota genetics, Gene Expression, Genetic Markers, Genetic Vectors, Genetics, Microbial methods, Integrases biosynthesis, Molecular Biology methods
- Abstract
Selectable marker recycling is a basic technique in bioengineering. However, this technique is usually unavailable in non-model microorganisms. In this study, we proposed a simple and efficient method for selectable marker recycling in the astaxanthin-synthesizing yeast Xanthophyllomyces dendrorhous. This method was based on a Cre-loxP system, in which the transient expression of the Cre recombinase was controlled by a genetically unstable vector independent of episomal plasmids and inducible promoters. The selectable markers in single-gene locus and multigene loci were removed along with the loss of the Cre vector with a ratio of 100% and 29%, respectively. The significance of the method was highlighted by the finding that stable autotrophic mutants were not readily obtained in X. dendrorhous. Comparative studies in X. dendrorhous and the non-homologous end joining dominant yeast Yarrowia lipolytica suggested that the method could be universally used in homologous recombination dominant yeasts.
- Published
- 2019
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