Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Isolation of a Promoter Element that is Functional in Bacillus subtilis for Heterologous Gene Expression

  • Maeng, Chang-Jae (Environmental Bioresources Lab., Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Hyung-Kwoun (Environmental Bioresources Lab., Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Sun-Yang (inBioNET Corporation) ;
  • Koo, Bon-Tag (inBioNET Corporation) ;
  • Oh, Tae-Kwang (Environmental Bioresources Lab., Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Jung-Kee (Environmental Bioresources Lab., Korea Research Institute of Bioscience and Biotechnology & inBioNET Corporation)
  • Published : 2001.02.01
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Abstract

To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

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