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KMID : 0545119910010010017
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Journal of Microbiology and Biotechnology
1991 Volume.1 No. 1 p.17 ~ p.21
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Cloning of a β-Xylosidase Gene from Alkalophilic Bacillus sp. and its Expression in Escherichia coli
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Yu, Ju Hyun
Kang, Yun Sook/Park, Young Seo
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Abstract
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A gene coding for (β-xylosidase in alkalophilic Bacillus sp. YC-335 isolated from soil was cloned into Escherichia coli HB101 using plasmid pBR322. The recombinant plasmid pYK40 was isolated, and the cloned HindIII fragment was 15 kilobases (kb). To reduce the size of the inserted DNA fragment of pYK40, the 15 kb HindIII fragment was subjected to a series of subclonings. A 6 kb subfragment was found to code for β-xylosidase activity, and the recombinant plasmid was named pYK44. Southern hybridization analysis revealed that the cloned gene hybridized with 3.5 kb, 1.5 kb, and 1.0 kb of HindIII cleaved chromosomal DNA from Bacillus sp. YC-335. (β-xylosidase activity produced by recombinant E. coli was found to be 11 times higher than that produced by Bacillus sp. YC-335. Xylan was required to induce the production of β-xylosidase in Bacillus sp. YC-335.
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