Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Identification of a Gene for Aerobic Growth with a SoxS Binding Sequence in Escherichia coli by Operon Fusion Techniques

  • Lee, Yong-Chan (Graduate School of Biotechnology, Korea University) ;
  • Kwon, Hyung-Bae (Graduate School of Biotechnology, Korea University) ;
  • Lee, Sang-Ho (Graduate School of Biotechnology, Korea University) ;
  • Kwon, Hye-Won (Graduate School of Biotechnology, Korea University) ;
  • Sung, Ha-Chin (Graduate School of Biotechnology, Korea University) ;
  • Kim, Joon (Graduate School of Biotechnology, Korea University) ;
  • Choe, Mu-Hyeon (Graduate School of Biotechnology, Korea University)
  • Published : 2001.12.01
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Abstract

Eight Escherichia coli cells with aerobic growth deflects were isolated by the insertion of ${\lambda}placMu53$, a hybrid bacteriophage of ${\lambda}$ and Mu, which created transcriptional fusion to lacZY. Two of these mutants, CLIO and CLl2, were irradiated with UV to obtain specialized transducing phages. The phages that took out the neighboring chromosomal DNA of the related gene responsible for deflective aerobic growth were identified. The in vivo cloned chromosomal sequence revealed that the mutated gene of CLIO was located at min 34.5 on the Escherichia coli linkage map and 1,599,515 on the physical map. The physical map indicated that there were 7 cistrons in the operon. We named this operon oxg10. The promoter sequence of oxg10 exhibited a possible binding site far SoxS, a transcriptional regulator that activates the transcription of various SoxRS regulon genes. Transferring the oxg10:: ${\lambda}placMu53$ mutation into the wild-type strain, RZ4500, resulted in the inhibition of normal aerobic growth, while the salute mutation in strain MO inhibited aerobic cell growth completely. The full operon sequences of oxg10 were cloned from the Excherichia coli genomic library. The mutated gene of CLl2 was identified to be a sucA gene encoding the ${\alpha}$-ketoglutarate dehydrogenase El component in the TCA cycle.

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