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Molecular Cloning and Expression of the $eta$-Xylosidase Gene (xylB) of Bacillus stearothermophilus in Escherichia coli
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  • Molecular Cloning and Expression of the $eta$-Xylosidase Gene (xylB) of Bacillus stearothermophilus in Escherichia coli
  • Molecular Cloning and Expression of the $eta$-Xylosidase Gene (xylB) of Bacillus stearothermophilus in Escherichia coli
저자명
Suh. Jung-Han,Eom. Soo-Jung,Cho. Ssang-Goo,Choi. Yong-Jin
간행물명
Journal of microbiology and biotechnology
권/호정보
1996년|6권 5호|pp.331-335 (5 pages)
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한국미생물생명공학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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The second $eta$-Xylosidase gene (xylB) from Bacillus stearothermophilus was isolated from the genomic library, cloned into pBR322, and subsequently transferred into Escherichia coli HB101. Six out of 10, 000 transformants were selected from the selective LB medium supplemented with p-nitrophenyl-$alpha$-L-arabinofuranoside (pNPAf) and ampicillin ($50mu g$/ml) based on their ability to form a yellow ring around the colony. One of the clones was found to harbor the recombinant plasmid with 5.0 kb foreign DNA, which was identical to the $alpha$-L-arabinofuranosidase gene (arfI) previously cloned in this lab, while the other five had 3.5 kb of the foreign DNA. Southern blotting experiments confirmed that the 3.5 kb insert DNA was from B. stearothermophilus chromosomal DNA. A zymogram with 4-methylumbelliferyl-$alpha$-L-arabinofuranoside as the enzyme substrate revealed that the cloned gene product was one of the mutiple $alpha$-L-arabinofuranosidases produced by B. stearothermophilus. Unlike the arfI gene product, the product of the gene on the insert DNA (xylB) showed an activity not only on pNPAf but also on oNPX suggesting that the cloned gene product could be a bifunctional enzyme having both $alpha$-L-arabinofuranosidase and $eta$-xylosidase activities.