caa a22 4500 606213295 CHVBK 20210128101046.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s10295-015-1684-8 doi (NATIONALLICENCE)springer-10.1007/s10295-015-1684-8 EasyClone 2.0: expanded toolkit of integrative vectors for stable gene expression in industrial Saccharomyces cerevisiae strains [Elektronische Daten] [Vratislav Stovicek, Gheorghe Borja, Jochen Forster, Irina Borodina] 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. The Author(s), 2015 Industrial yeast nationallicence Integrative vectors nationallicence Heterologous gene expression nationallicence Metabolic engineering nationallicence Xylose utilization nationallicence Stovicek Vratislav The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970, Hørsholm, Denmark aut Borja Gheorghe The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970, Hørsholm, Denmark aut Forster Jochen The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970, Hørsholm, Denmark aut Borodina Irina The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970, Hørsholm, Denmark aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/11(2015-11-01), 1519-1531 1367-5435 42:11<1519 2015 42 10295 https://doi.org/10.1007/s10295-015-1684-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1684-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Stovicek Vratislav The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970, Hørsholm, Denmark aut NATIONALLICENCE 700 1- Borja Gheorghe The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970, Hørsholm, Denmark aut NATIONALLICENCE 700 1- Forster Jochen The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970, Hørsholm, Denmark aut NATIONALLICENCE 700 1- Borodina Irina The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970, Hørsholm, Denmark aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/11(2015-11-01), 1519-1531 1367-5435 42:11<1519 2015 42 10295