Cloning, sequencing and disruption of theARG8 gene encoding acetylornithine aminotransferase in the petite-negative yeast Kluyveromyces lactis
✍ Scribed by Janssen, Antje; Chen, Xin Jie
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 154 KB
- Volume
- 14
- Category
- Article
- ISSN
- 0749-503X
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✦ Synopsis
A recombinant plasmid was isolated from a Kluyveromyces lactis genomic DNA library which complements a Saccharomyces cerevisiae arg8 mutant defective in the gene encoding acetylornithine aminotransferase. The complementation activity was found to reside within a 2.0 kb DNA fragment. Nucleotide sequence analysis revealed an open reading frame able to encode a 423-residue protein sharing 68.1% and 35.0% sequence identities with the products of the ARG8 and argD genes of S. cerevisiae and Escherichia coli. That the cloned gene, KlARG8, is the functional equivalent of S. cerevisiae ARG8 was supported by a gene disruption experiment which showed that K. lactis strains carrying a deleted chromosomal copy of KlARG8 are auxotrophic for arginine.