Work

Ty retrotransposon element based multiple integration toolkit for Saccharomyces cerevisiae

Year: 2025

Type: article

Abstract: Extra-high-level overexpression of single or multiple specific proteins by integrating specific genes in the genome is vital to achieve the stable and efficient production of target proteins and metabolites in S. cerevisiae . Five families of Ty elements in the genome of S. cerevisiae CEN.PK2-1D, which could have dozens to hundreds of copies, have been employed to achieve massive gene expression. By engineering nine selective markers, six of them ( TRP1 , LEU2 , URA3 , HIS5 , natMX and hphMX ) achieve stably high copy integration (>15 copies) at Ty sites. Fluorescence proteins and taxifolin biosynthesis pathway genes were overexpressed to verify the toolkit. The titer of protein phiYFP in the multiple integration strain reached 1.6 g/L (268.1 mg/g DCW), and its fluorescence intensity was 3.3 times higher than that in the episomal overexpression strain. For taxifolin biosynthesis, 14 genes were integrated into three different Ty sites using three selective markers from the toolkit, resulting in 277.6 mg/L taxifolin accumulation from glucose.

Source: Synthetic and Systems Biotechnology

DOI: 10.1016/j.synbio.2025.04.011

Authors: Song Gao, Weizhu Zeng, Dong Li, Sha Xu, Jingwen Zhou

Institution: Jiangnan University

Language: English

Topic: Fungal and yeast genetics research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,500

Funders: Innovative Research Group Project of the National Natural Science Foundation of China, National Natural Science Foundation of China, National Key Research and Development Program of China