Vectors for gene expression and amplification in the yeast Yarrowia lipolytica

Emese Vissi, Josep Clotet, Eulalia De Nadal, Anna Barceló, Éva Bakó, Pl Gergely, Viktor Dombrdi, Joaquín Ariño

Research output: Contribution to journalArticleResearchpeer-review

106 Citations (Scopus)

Abstract

New vector systems were developed for gene expression in Y. lipolytica. These plasmids contain: (a) as integration target sequences, either a rDNA region or the long terminal repeat zeta of the Y. lipolytica retrotransposon Ylt1; (b) the YlURA3 gene as selection marker for Y. lipolytica, either as the non-defective ura3d1 allele for single integration or the promotor truncated ura3d4 allele for multiple integration; (c) the inducible ICL1 or XPR2 promoters for gene expression; and (d) unique restriction sites for gene insertion. Multiple plasmid integration occurred as inserted tandem-repeats, which are present at 3-39 copies per cell. A correlation between gene copy number and the expressed enzyme activity was demonstrated with Escherichia coli lacZ as reporter gene under the control of the regulated ICL1 promoter. Increases in copy numbers from 5 to 13 for the lacZ expression cassettes resulted in an up to 10-11-fold linear increase of the β-galactosidase activity in multicopy transformants during their growth on ethanol or glucose, compared with the low-copy replicative plasmid transformants (1.6 plasmid copies). These new tools will enhance the interest in Y. lipolytica as an alternative host for heterologous protein production. Copyright © 2000 John Wiley & Sons, Ltd.
Original languageEnglish
Pages (from-to)97-113
JournalYeast
Volume18
Issue number2
DOIs
Publication statusPublished - 30 Jan 2001

Keywords

  • Gene amplification
  • Gene expression
  • Integrative vectors
  • LTR zeta
  • rDNA
  • Yarrowia lipolytica
  • Yeast

Fingerprint Dive into the research topics of 'Vectors for gene expression and amplification in the yeast Yarrowia lipolytica'. Together they form a unique fingerprint.

  • Cite this

    Vissi, E., Clotet, J., De Nadal, E., Barceló, A., Bakó, É., Gergely, P., Dombrdi, V., & Ariño, J. (2001). Vectors for gene expression and amplification in the yeast Yarrowia lipolytica. Yeast, 18(2), 97-113. https://doi.org/10.1002/1097-0061(20010130)18:2<97::AID-YEA652>3.0.CO;2-U