Generation of a stable non-reverting Leu- mutant of Kluyveromyces lactis by gene disruption.
The LEU2 gene coding for beta-isopropylmalate dehydrogenase of the yeast Kluyveromyces lactis strain AWJ137 was disrupted. In the resulting Leu- strain a 0.57 x 10(3)-base pairs PstI/BglII fragment of the LEU2 coding region was replaced by the TRP1 gene of Saccharomyces cerevisiae. The mutant strain was characterized by stability tests and a physical map of the disrupted region was established by restriction-enzyme analysis combined with hybridization experiments. The usefulness of the mutant strain as a recipient was shown by transformation experiments.
Meinhardt
F
F
Institut für Mikrobiologie, Westfälische Wilhelms-Universität Münster, Germany.
Holtwick
R
R
D050539:3-Isopropylmalate Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000235:genetics*; D001483:Base Sequence; D001709:Biotechnology; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D007716:Kluyveromyces / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D015183:Restriction Mapping; D014170:Transformation, Genetic
D004271:DNA, Fungal; D000429:Alcohol Oxidoreductases; D050539:3-Isopropylmalate Dehydrogenase