Establishment of genomic library technology mediated by non-homologous end joining mechanism in Yarrowia lipolytica.
Yarrowia lipolytica
genomic library
non-homologous end joining
synthetic biology
Journal
Science China. Life sciences
ISSN: 1869-1889
Titre abrégé: Sci China Life Sci
Pays: China
ID NLM: 101529880
Informations de publication
Date de publication:
12 2021
12 2021
Historique:
received:
16
11
2020
accepted:
28
01
2021
pubmed:
5
3
2021
medline:
19
3
2022
entrez:
4
3
2021
Statut:
ppublish
Résumé
Genomic variants libraries are conducive to obtain dominant strains with desirable phenotypic traits. The non-homologous end joining (NHEJ), which enables foreign DNA fragments to be randomly integrated into different chromosomal sites, shows prominent capability in genomic libraries construction. In this study, we established an efficient NHEJ-mediated genomic library technology in Yarrowia lipolytica through regulation of NHEJ repair process, employment of defective Ura marker and optimization of iterative transformations, which enhanced genes integration efficiency by 4.67, 22.74 and 1.87 times, respectively. We further applied this technology to create high lycopene producing strains by multi-integration of heterologous genes of CrtE, CrtB and CrtI, with 23.8 times higher production than rDNA integration through homologous recombination (HR). The NHEJ-mediated genomic library technology also achieved random and scattered integration of loxP and vox sites, with the copy number up to 65 and 53, respectively, creating potential for further application of recombinase mediated genome rearrangement in Y. lipolytica. This work provides a high-efficient NHEJ-mediated genomic library technology, which enables random and scattered genomic integration of multiple heterologous fragments and rapid generation of diverse strains with superior phenotypes within 96 h. This novel technology also lays an excellent foundation for the development of other genetic technologies in Y. lipolytica.
Identifiants
pubmed: 33660223
doi: 10.1007/s11427-020-1885-x
pii: 10.1007/s11427-020-1885-x
doi:
Substances chimiques
Lycopene
SB0N2N0WV6
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
2114-2128Informations de copyright
© 2021. Science China Press and Springer-Verlag GmbH Germany, part of Springer Nature.
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