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Abstract
Metschnikowia pulcherrima is a non-conventional yeast with the potential to be used in bi-otechnological processes, especially involving low-cost feedstock exploitation. However, there are a lack of tools for researching it at a molecular level and for producing genetically modified strains. We tested the amenability to genetic modification of ten different strains, establishing a transformation protocol based on LiAc/PEG that allows us to introduce heterologous DNA. Non-homolo-gous integration was broadly successful and homologous recombination was successful in two strains. Chemical inhibition of non-homologous end joining recombination had a modest effect on the improvement of homologous recombination rates. Removal of selective markers via flippase recombinase was successful across integrated loci except for those targeted to the native URA3 lo-cus, suggesting that the genome sequence or structure alters the efficacy of this system.
Original language | English |
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Article number | 290 |
Pages (from-to) | 1-11 |
Number of pages | 11 |
Journal | Microorganisms |
Volume | 9 |
Issue number | 2 |
DOIs | |
Publication status | Published - 31 Jan 2021 |
Keywords
- Biotechnology
- Homologous recombination
- Nonconventional yeasts
ASJC Scopus subject areas
- Microbiology
- Virology
- Microbiology (medical)
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Dive into the research topics of 'Variation among metschnikowia pulcherrima isolates for genetic modification and homologous recombination'. Together they form a unique fingerprint.Projects
- 1 Finished
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Integrated Energy Efficient Microwave and Unique Fermentation Processes for Pilot Scale Production of High Value Chemical from Lignocellulosic Waste
Chuck, C., Henk, D., Leak, D., McManus, M. & Scott, R.
Engineering and Physical Sciences Research Council
1/03/16 → 31/01/21
Project: Research council