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DNA Double-Strand Break-Induced Gene Amplification in Yeast

Publikation: Bidrag til bog/antologi/rapportBidrag til bog/antologi

Precise control of the gene copy number in the model yeast Saccharomyces cerevisiae may facilitate elucidation of enzyme functions or, in cell factory design, can be used to optimize production of proteins and metabolites. Currently, available methods can provide high gene-expression levels but fail to achieve accurate gene dosage. Moreover, strains generated using these methods often suffer from genetic instability resulting in loss of gene copies during prolonged cultivation. Here we present a method, CASCADE, which enables construction of strains with defined gene copy number. With our present system, gene(s) of interest can be amplified up to nine copies, but the upper copy limit of the system can be expanded. Importantly, the resulting strains can be stably propagated in selection-free media.

OriginalsprogEngelsk
TitelHomologous Recombination : Methods and Protocols
RedaktørerAndrés Aguilera, Aura Carreira
Antal sider14
ForlagHumana Press
Publikationsdato2021
Sider239-252
Kapitel17
ISBN (Trykt)978-1-0716-0643-8
ISBN (Elektronisk)978-1-0716-0644-5
DOI
StatusUdgivet - 2021
NavnMethods in Molecular Biology
Vol/bind2153
ISSN1064-3745

ID: 249857300