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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Cloning, Sequencing and Expression of dTDP-D-Glucose 4,6-Dehydratase Gene from Streptomyces antibioticus $T\ddot{u}99$, a Producer of Chlorothricin

  • Received : 1995.11.24
  • Published : 1996.05.31
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Abstract

DNA fragments, homologous to the dTDP-D-glucose 4,6-dehydratase gene, obtained from the genomic DNA of Streptomyces antibioticus $T\ddot{u}99$, a producer of the unusual macrolide antibiotic chlorothricin, were cloned and sequenced. This dehydratase gene was designated as oxil. The coding region of the oxil gene is composed of 987 bp, and analysis of the DNA sequence data reveals sequences for the gene products of 329 amino acids (molecular weight of 36,037). The deduced amino acids are 59% identical to the StrE, dTDP-D-glucose 4,6-dehydratase from the streptomycin pathway. The oxil's function was examined by expressing it in E. coli using the T7 RNA polymerase/promoter system (pRSET) to produce an active fusion protein including a his tag. This enzyme shows specificity of substrate, specific only to dTDP-D-glucose.

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