Inactivation of Yarrowia lipolytica YlACL2 gene Coding Subunit of ATP Citrate Lyase Using CRISPR/Cas9 System

Autor:	M. O. Taratynova, D. A. Egorova, Evgeniya Y. Yuzbasheva, A. I. Solovyev, Iuliia M. Kosikhina, E. B. Vinogradova, D. A. Dementev, S. P. Sineoky, Tigran V. Yuzbashev
Rok vydání:	2020
Předmět:	ATP citrate lyase biology Cas9 Protein subunit Uracil Yarrowia biology.organism_classification Applied Microbiology and Biotechnology Biochemistry chemistry.chemical_compound chemistry CRISPR Homologous recombination Gene
Zdroj:	Applied Biochemistry and Microbiology. 56:885-892
ISSN:	1608-3024 0003-6838
DOI:	10.1134/s0003683820090112
Popis:	In this study, YlACL2 was inactivated by two methods: traditional approach based on homologous recombination and uracil marker and markerless system using CRISPR/Cas9. The efficiency of YlACL2 inactivation using traditional approach was 4% (one ΔYlacl2 strain out of 24 tested transformants) whereas knockout efficiency using CRISPR/Cas9 system was 75% (18 ΔYlacl2 strains out of 24 tested transformants). YlACL2 null mutant strains were not able to utilize citrate as a single carbon source. Growth kinetics was investigated in the media with glucose and acetate as a single carbon source. The fact that ΔYlacl2 is able to grow in the minimal medium with glucose as a single carbon source provides evidence that there is an alternative source of acetyl-CoA on carbohydrate substrates in Y. lipolytica.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::9c163ce82d41d46f825ed392dec90ae8 https://doi.org/10.1134/s0003683820090112 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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