BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Cloning and Characterization of the Mycobacterium bovis BCG panB Gene Encoding Ketopantoate Hydroxymethyltransferase

  • Kim, Jin-Koo (Cell Biology Laboratory, Korea Research Institute of Bioscience & Biotechnology (KRIBB)) ;
  • Kim, Kwang-Dong (Cell Biology Laboratory, Korea Research Institute of Bioscience & Biotechnology (KRIBB)) ;
  • Lim, Jong-Seok (Cell Biology Laboratory, Korea Research Institute of Bioscience & Biotechnology (KRIBB)) ;
  • Lee, Hee-Gu (Cell Biology Laboratory, Korea Research Institute of Bioscience & Biotechnology (KRIBB)) ;
  • Kim, Sang-Jae (Korea Institute of Tuberculosis, KNTA) ;
  • Cho, Sang-Hyun (Korea Institute of Tuberculosis, KNTA) ;
  • Jeong, Won-Hwa (Boryung Biophanna Co.) ;
  • Choe, In-Seong (Cell Biology Laboratory, Korea Research Institute of Bioscience & Biotechnology (KRIBB)) ;
  • Chung, Thi-Wha (Cell Biology Laboratory, Korea Research Institute of Bioscience & Biotechnology (KRIBB)) ;
  • Paik, Sang-Gi (Department of Biology, Chungnam National University) ;
  • Choe, Yong-Kyung (Cell Biology Laboratory, Korea Research Institute of Bioscience & Biotechnology (KRIBB))
  • Received : 2001.04.20
  • Accepted : 2001.05.14
  • Published : 2001.07.31
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Abstract

The Mycobacterium bovis BCG panB gene, encoding ketopantoate hydroxymethyltransferase (KPHMT), was cloned from a ${\lambda}gt11$ genomic library and sequenced. The DNA sequence encodes a protein that contains 281 amino acid residues (M, 29,337) with a high similarity to the KPHMTs. Subcloning of a 846 by open reading frame (ORF), but not a 735 by ORF, into the vector pUC19 led to complementation of the panB mutant of Escherichia coli. The BCG pang gene was overexpressed in E. coli and the KPHMT purified to homogeneity The recombinant protein was further confirmed by an enzymatic assay.

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