Summary
A Yarrowia lipolytica yeast gene bank has been constructed in E. coli in an integrative plasmid vector containing the homologous LEU2 gene, and used to transform a leu2 lys5 yeast strain. The LYS5 gene encoding saccharopine dehydrogenase (SDH) has been rescued in E. coli from the chromosome of prototrophic transformants, in which the hybrid plasmid had integrated at the leu2 locus. Evidence that the rescued clone contains the LYS5 gene comes from complementation tests, genetic crosses, and SDH assay. Further characterization of the gene has been achieved by deletion mapping and subcloning, and by demonstrating the presence of transcripts hybridizing to this sequence.
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Xuan, JW., Fournier, P. & Gaillardin, C. Cloning of the LYS5 gene encoding saccharopine dehydrogenase from the yeast Yarrowia lipolytica by target integration. Curr Genet 14, 15–21 (1988). https://doi.org/10.1007/BF00405848
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DOI: https://doi.org/10.1007/BF00405848