Journal article
EasyClone 2.0: expanded toolkit of integrative vectors for stable gene expression in industrial Saccharomyces cerevisiae strains
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Applied Metabolic Engineering, Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
Yeast Metabolic Engineering, Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark4
Yeast Cell Factories, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
Saccharomyces cerevisiae is one of the key cell factories for production of chemicals and active pharmaceuticals. For large-scale fermentations, particularly in biorefinery applications, it is desirable to use stress-tolerant industrial strains. However, such strains are less amenable for metabolic engineering than the standard laboratory strains.
To enable easy delivery and overexpression of genes in a wide range of industrial S. cerevisiae strains, we constructed a set of integrative vectors with long homology arms and dominant selection markers. The vectors integrate into previously validated chromosomal locations via double cross-over and result in homogenous stable expression of the integrated genes, as shown for several unrelated industrial strains.
Cre-mediated marker rescue is possible for removing markers positioned on different chromosomes. To demonstrate the applicability of the presented vector set for metabolic engineering of industrial yeast, we constructed xylose-utilizing strains overexpressing xylose isomerase, xylose transporter and five genes of the pentose phosphate pathway.
| Language: | English |
|---|---|
| Publisher: | Springer Berlin Heidelberg |
| Year: | 2015 |
| Pages: | 1519-1531 |
| Journal subtitle: | Official Journal of the Society for Industrial Microbiology and Biotechnology |
| ISSN: | 14765535 and 13675435 |
| Types: | Journal article |
| DOI: | 10.1007/s10295-015-1684-8 |
| ORCIDs: | Stovicek, Vratislav , Borja Zamfir, Gheorghe Manuel , Förster, Jochen and Borodina, Irina |
Heterologous gene expression Industrial yeast Integrative vectors Metabolic engineering Xylose utilization
Aldose-Ketose Isomerases Biochemistry, general Bioinformatics Biomedical and Life Sciences Biotechnology Chromosomes, Fungal Crossing Over, Genetic Fermentation Gene Expression Regulation, Fungal Genetic Engineering Genetic Markers Genetic Vectors Inorganic Chemistry Life Sciences Metabolic Engineering Microbiology Pentose Phosphate Pathway Saccharomyces cerevisiae Xylose xylose isomerase
