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Food science and biotechnology v.19 no.3, 2010년, pp.849 - 855   SCIE
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Molecular Cloning of the Genes for GDP-mannose 4, 6-dehydratase and GDP-L-fucose Synthetase from Bacteroides thetaiotaomicron

Jang, Mi-Hee    (School of and Biotechnology and Bioengineering, and Institute of Bioscience and Biotechnology, Kangwon National University   ); Lee, Won-Heong    (Department of Agricultural Biotechnology, Seoul National University   ); Shin, So-Yeon    (Department of Agricultural Biotechnology, Seoul National University   ); Han, Nam-Soo    (Department of Food Science and Technology, Chungbuk National University   ); Seo, Jin-Ho    (Department of Agricultural Biotechnology, Seoul National University   ); Kim, Myoung-Dong    (School of and Biotechnology and Bioengineering, and Institute of Bioscience and Biotechnology, Kangwon National University  );
  • 초록

    Genes encoding GDP-mannose 4, 6-dehydratase (GMD) and GDP-L-fucose synthetase (GFS) were cloned from Bacteroides thetaiotaomicron and overexpressed in recombinant Escherichia coli by constructing isopropyl- $\beta$ -D-thiogalactopyranoside (IPTG)-inducible expression vectors. GMD and GFS genes from B. thetaiotaomicron were 60 and 45%, respectively, identical to those from E. coli K12 over their entire lengths. An optimum expression condition of $30^{\circ}C$ and 0.1 mM IPTG was chosen for maximum soluble expression of B. thetaiotaomicron GMD and GFS in recombinant E. coli BL21(DE3). Functional expression of B. thetaiotaomicron GMD and GFS in recombinant E. coli strains was confirmed by measuring intracellular GDP-L-fucose content.


  • 주제어

    GDP-L-fucose .   GDP-mannose 4,6-dehydratase .   GDP-L-fucose synthetase .   Bacteroides thetaiotaomicron .   recombinant Escherichia coli.  

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