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Current Genetics

, Volume 11, Issue 5, pp 369–375 | Cite as

LEU2 directed expression of β-galactosidase activity and phleomycin resistance in Yarrowia lipolytica

  • Claude Gaillardin
  • Anne-Marie Ribet
Original Articles

Summary

The nucleotide sequence of a 968 by DNA fragment spanning the promoter and the 5′ upstream sequence of the LEU2 coding sequence of the yeast Yarrowia lipolytica has been determined. A LEU2:lacZ gene fusion has been constructed and expressed in transformed yeast cells, showing that as few as 232 by of the LEU2 promotor were sufficient to direct gene expression. In order to develop new markers for transformation of this yeast, the LEU2 initiation codon was destroyed by in vitro mutagenesis and replaced by a cloning site. A gene confering phleomycin resistance in E. coli was attached to the LEU2 promoter and shown to be efficiently expressed in yeast: direct selection of phleomycin resistant transformants was possible.

Key words

Yarrowia lipolytica LEU2 promoter β-galactosidase Phleomycin resistance 

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Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • Claude Gaillardin
    • 1
  • Anne-Marie Ribet
    • 1
  1. 1.Institut National AgronomiqueLaboratoire de GénétiqueParis Cédex 05France

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