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The Yarrowia lipolytica LEU2 gene
L S Davidow1, F S Kaczmarek, J R DeZeeuw
1Pfizer Central Research, Groton, CT 06340.
Current Genetics
|January 1, 1987
Summary
Researchers sequenced the beta-isopropylmalate dehydrogenase gene (LEU2) in Yarrowia lipolytica, finding a 405-codon open reading frame. This yeast LEU2 gene shows significant homology to Saccharomyces cerevisiae, aiding in understanding yeast metabolic pathways.
Area of Science:
- Molecular Biology
- Yeast Genetics
Background:
- The beta-isopropylmalate dehydrogenase (LEU2) gene is crucial for leucine biosynthesis in yeast.
- Understanding gene structure and regulation in dimorphic yeast like Yarrowia lipolytica is important for biotechnological applications.
Purpose of the Study:
- To sequence and characterize the LEU2 gene from Yarrowia lipolytica.
- To compare its genetic and protein sequence with the homologous gene in Saccharomyces cerevisiae.
Main Methods:
- DNA sequencing of a 2810 bp fragment.
- Bioinformatic analysis of open reading frame (ORF) and flanking regions.
- Homology comparison at both DNA and protein levels.
Main Results:
- A 2810 bp DNA fragment containing the Y. lipolytica LEU2 gene was sequenced.
- An ORF of 405 codons was identified, predicting a 43,366 MW protein.
- 64% protein and 61% DNA sequence homology was found with S. cerevisiae LEU2.
- Shared general structural features in flanking regions were observed, including regulatory elements like TATA and CAAT boxes.
- Transcription initiates approximately 40 bp upstream of the ATG start codon, producing a 1.4 kb mRNA.
Conclusions:
- The Y. lipolytica LEU2 gene is structurally similar to its S. cerevisiae counterpart.
- Sequence homology suggests functional conservation of the beta-isopropylmalate dehydrogenase enzyme.
- Characterization provides insights into gene regulation and potential use in metabolic engineering of Y. lipolytica.