EasyClone 2.0: expanded toolkit of integrative vectors for stable gene expression in industrial Saccharomyces cerevisiae strains

Vratislav Stovicek, Gheorghe Manuel Borja Zamfir, Jochen Förster, Irina Borodina

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Abstract

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.
Original languageEnglish
JournalJournal of Industrial Microbiology and Biotechnology
Volume42
Issue number11
Pages (from-to)1519-1531
Number of pages13
ISSN1367-5435
DOIs
Publication statusPublished - Oct 2015

Bibliographical note

This article is published with open access at Springerlink.com

Keywords

  • Industrial yeast
  • Integrative vectors
  • Heterologous gene expression
  • Metabolic engineering
  • Xylose utilization

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