Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Development of an E. coli Expression Cassette for the Efficient Production of a Harmful Protein

  • Kim Ok Soo (Major in Bioscience and Biotechnology, Division of Environmental Engineering and Biotechnology, Silla University) ;
  • Kwak Hwan Jong (Department of Pathology, College of Medicine, The Catholic University of Korea) ;
  • Lee Jae-Hwa (Major in Bioscience and Biotechnology, Division of Environmental Engineering and Biotechnology, Silla University) ;
  • Ha Jong Myung (Major in Bioscience and Biotechnology, Division of Environmental Engineering and Biotechnology, Silla University) ;
  • Ha Bae-Jin (Major in Bioscience and Biotechnology, Division of Environmental Engineering and Biotechnology, Silla University) ;
  • Lee Sang-Hyeon (Major in Bioscience and Biotechnology, Division of Environmental Engineering and Biotechnology, Silla University)
  • Published : 2004.10.01
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Abstract

In order to produce a harmful protein more efficiently, this expression cassette, dubbed pCol-MICT, is directed by the colicin promoter, and was constructed by the insertion of a $rrnBT_1T_2$ fragment of pEXP7, and a MxelnteinCBD fragment of pTXB3, into pSH375. To test whether harmful proteins, including proteolytic enzymes, could be effectively produced by this cassette, the carboxypeptidase (CPase) Taq gene was inserted into the pCol-MICT cassette to yield pCol-CPase Taq-MICT. E coli W3l 10 tells harboring pCol-CPase Taq-MICT produced a large quantity of this enzyme, as much as 47.2 mg of purified from per liter of culture, when cultured in the presence of mitomycin C ($0.4{\mu}g/mL$). This indicates that the colicin promoter-controlled E, coli expression cassette was able to produce almost 8 times of protein than the conventional tar promoter-based system, and that this cassette may be useful in the Synthesis of other harmful proteins.

Keywords

References

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