Cloning and nucleotide sequence analysis of xylose reductase (XR) gene from thermotolerant methylotrophic yeast Ogataea siamensis N22
DOI:
https://doi.org/10.14456/tjg.2009.8Keywords:
xylose reductase, gene cloning, Ogataea siamensis, thermotolerant yeastAbstract
The xylose reductase (XR) gene from thermotolerant methylotrophic yeast isolated in Thailand, Ogataea siamensis N22, was isolated using specific primers designed from the conserved regions of various yeast XR genes. A partial XR gene of O. siamensis N22, was obtained as a 516 bp PCR product and the deduced amino acid sequence showed 74% identity to aldose reductase gene of Candida boidinii and and 67% identity to xylose reductase gene of Kluyveromyces lactis and Pichia guilliermondii. The 1-3 kb fragments of digested genomic DNA of O. siamensis N22 was cloned and transformed into Escherichia coli. The recombinant plasmid containing the partial nucleotide sequence of XR gene was obtained using PCR screening. Nucleotide sequence analysis of the partial O. siamensis N22 XR gene revealed 678 bp encoding 226 amino acids which showed 72% identity to aldose reductase from Candida boidinii and 69% identity to xylose reductase from Pichia guilliermondii. This is the first report on xylose reductase gene of O. siamensis N22.
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