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KMID : 0613820020120020188
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Journal of Life Science
2002 Volume.12 No. 2 p.188 ~ p.199
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Molecular Cloning and Analysis of Nucleotide Sequence of Xylanase Gene (xynK) from Bacillus pumilus TX703
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Park Young-Seo
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Abstract
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A gene coding for xylanase from thermo-tolerant Bacillus pumilus TX703 was cloned into Escherichia coli DH5α using pUC19. Among 7,400 transformants, four transformants showed clear zones on the detection agar plates containing oat-spelts xylan. One of them which showed highest xylanase activity was selected and its recombinant plasmid, named pXES106, was found to carry 2.24 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynK) was determined, xynK gene was found to consist of 1,227 base-pair open reading frame coding for a polypeptide of 409 amino acids with a deduced molecular weight of 48 kDa. The coding sequence was preceded by a putative ribosome binding site, the transcription initiation signals, and cis-acting catabolite responsive element. The deduced amino acids sequence of xylanase is similar to those of the xylanases from Hordeum vulgare (barley) and Clostridium thermocellum, with 39 and 31% identical residues, respectively. The amino acids sequence of this xylanase was quite different from those of the xylanases from other Bacillus species.
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KEYWORD
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xylanase gene, cloning, nucleotide sequence, Bacillus pumilus
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