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Summary: A mutant of Hansenula polymorpha defective in β-isopropylmalate dehydrogenase was isolated. It was transformed with YEp13, a shuttle vector containing the Saccharomyces cerevisiae LEU2 gene, either using the protoplasting method or by treatment with LiCl. Leu+ transformants were recovered at a frequency of 30-60 μg DNA)−1. The transformants were shown to contain an autonomously replicating plasmid by stability tests, hybridization experiments and by the recovery of YEp13 from transformed cells via transformation of Escherichia coli. YEp13 linearized by BamHI digestion was also effective in transformation; the linearized plasmid was shown to recircularize and replicate autonomously in the transformants.
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