Molecular Cloning of the <i>leuB</i> Gene from <i>Bacteroides fragilis</i> by Functional Complementation in <i>Escherichia coli</i>

J-STAGE
  • Sarker Mahfuzur R.
    Department of Bacteriology, School of Medicine, The University of Tokushima
  • Akimoto Shigeru
    Department of Bacteriology, School of Medicine, The University of Tokushima
  • Ono Tsuneko
    Department of Bacteriology, School of Medicine, The University of Tokushima
  • Kinouchi Takemi
    Department of Bacteriology, School of Medicine, The University of Tokushima
  • Ohnishi Yoshinari
    Department of Bacteriology, School of Medicine, The University of Tokushima

抄録

Clones containing the Bacteroides fragilis leuB-complementing gene were isolated by screening of a B. fragilis genomic library constructed in Escherichia coli. One recombinant clone, designated pOT865, with the smallest DNA insert (4.5kb) could complement three independent leuB mutations in E. coli and the leuB-complementing determinant in pOT865 was localized to a region of 1.5-kb DNA. The results of Southern blot analysis suggested that a single copy of the cloned gene was present in the B. fragilis genome. The cloned fragment appeared to contain a sequence that could function as a promoter in E. coli and direct the synthesis of a 42-kDa protein. These results suggest that the cloned segment contains the structural gene for β-isopropylmalate dehydrogenase (leuB).

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