Title |
EasyClone 2.0: expanded toolkit of integrative vectors for stable gene expression in industrial Saccharomyces cerevisiae strains
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Published in |
Journal of Industrial Microbiology & Biotechnology, November 2015
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DOI | 10.1007/s10295-015-1684-8 |
Pubmed ID | |
Authors |
Vratislav Stovicek, Gheorghe M Borja, Jochen Forster, Irina Borodina |
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. |
X Demographics
Geographical breakdown
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France | 1 | 100% |
Demographic breakdown
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Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
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Denmark | 1 | <1% |
Germany | 1 | <1% |
Brazil | 1 | <1% |
Unknown | 234 | 99% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 42 | 18% |
Researcher | 41 | 17% |
Student > Master | 34 | 14% |
Student > Bachelor | 24 | 10% |
Student > Doctoral Student | 10 | 4% |
Other | 30 | 13% |
Unknown | 56 | 24% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 80 | 34% |
Agricultural and Biological Sciences | 69 | 29% |
Engineering | 4 | 2% |
Chemistry | 3 | 1% |
Medicine and Dentistry | 3 | 1% |
Other | 15 | 6% |
Unknown | 63 | 27% |