본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

Biotechnology and bioprocess engineering v.9 no.5, 2004년, pp.389 - 392   피인용횟수: 2

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  );
  • 초록

    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.


  • 주제어

    colicin promoter .   expression cassette .   E.   coli W3110 cells .   carboxypeptidase Taq .   mitomytin C.  

  • 참고문헌 (14)

    1. Hong, J.-K., M.-S. Park, F M. Raushel, and Y-H. Khang (2003) Expression and characterization of Escherichia coli prolidase with organophosphorus compounds. Biotechnol. Bioprocess Eng. 8: 126-129 
    2. Lee, S.-H., H. Taguchi, E. Yoshimura, E. Minagawa, S. Kaminogawa, T. Ohta, and H. Matsuzawa (1994) Carboxypeptidase Taq, a thermostable zinc enzyme, from Therrnus aquaticus YT-1: Molecular cloning, sequencing, and expression of the encoding gene in Escherichia coli. Biosci. Biotech. Biochem. 58: 1490-1495 
    3. Ebina, Y., Y. Takahara, K. Shirabe, M. Yamada, T. Nakazawa, and A. Nakazawa (1983) Plasmid-encoded regulation of colicin El gene expression. J. Bacteriol. 156: 487-492 
    4. Parker, R. C. (1986) Mitomycin C-induced bidirectional transcription from the colicin E1 promoter region in plasmid ColE1. Biochirn. Biophys. Acta 868: 39-44 
    5. Petit, M. A, W. Bedale, J. Osipiuk, C. Lu, M. Rajagopalan, P. McInerney, M. F Goodman, H. Echols (1994) Sequential folding of UmuC by the Hsp70 and Hsp60 chaperone complexes of Escherichia coli. J. Bioi. Chem. 269: 23824-23829 
    6. Sanger, F., S. Nicklen, and A. R. Coulson (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463-5467 
    7. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 
    8. Masaki, H., A Akutsu, T. Uozumi, and T. Ohta (1991) Identification of a unique specificity determinant of the colicin E3 immunity protein. Gene 107: 133-138 
    9. Rosen, H. (1957) A modified ninhydrin colorimetric analysis for amino acids. Arch. Biochem. Biophys. 67: 10-15 
    10. Masaki, H. and T. Ohta (1985) Colicin E3 and its immunity genes. J. Mol. Biol. 182: 217-227 
    11. Waleh, N. S., I. Sohel, J. B. Lazar, D. V. Hudson, P. Sze, P. A. Underhill, and P. H. Johnson (1992) Expression of synthetic genes encoding fused proteins under tight control of modified regulatory regions of the colicin operon. Gene 117: 7-14 
    12. Cavard, D., S. P. Howard, R. Lloubes, and C. Lazdunski (1989) High-level expression of the colicin A lysis protein. Mol. Gen. Genet. 217: 511-519 
    13. Waleh, N. S. and P. H. Johnson (1985) Structural and functional organization of the colicin E1 operon. Proc. Natl. Acad. Sci. USA 82: 8389-8393 
    14. Van den Elzen, P. J., M. J. Maat, H. H. B., Walters, E. Veltkamp, and H. J. Nijkamp (1982) The nucleotide sequence of the bacteriocin promoters of plasmids CIo DF13 and Col E1: Role of lexA repressor and cAmp in the regulation of promoter activity. Nucleic Acids Res. 10: 1913-1928 
  • 이 논문을 인용한 문헌 (2)

    1. 2008. "" Biotechnology and bioprocess engineering, 13(4): 470~475     
    2. Xu, Ming-Lu ; Wang, Lan ; Xu, Gui-Fang ; Wang, Myeong-Hyeon 2011. "Antidiabetes and Angiotensin Converting Enzyme Inhibitory Activity of Sonchus asper (L) Hill Extract" 생약학회지, 42(1): 61~67     

 저자의 다른 논문

  • 김옥수 (3)

    1. 2004 "Leucocin A로 형질전환된 효모의 항균 활성도" 한국생물공학회지 = Korean journal of biotechnology and bioengineering 19 (4): 291~294    
    2. 2005 "생쥐 도파민 수용쳬 조절인자 (DRRF) 유전자의 전사조절" 생명과학회지 = Journal of life science 15 (1): 55~60    
  • Kwak Hwan Jong (2)

  • 이재화 (88)

  • 하종명 (45)

  • Ha, Bae-Jin (77)

  • 이상현 (95)

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역

원문보기

무료다운로드
  • NDSL :
유료다운로드

유료 다운로드의 경우 해당 사이트의 정책에 따라 신규 회원가입, 로그인, 유료 구매 등이 필요할 수 있습니다. 해당 사이트에서 발생하는 귀하의 모든 정보활동은 NDSL의 서비스 정책과 무관합니다.

원문복사신청을 하시면, 일부 해외 인쇄학술지의 경우 외국학술지지원센터(FRIC)에서
무료 원문복사 서비스를 제공합니다.

NDSL에서는 해당 원문을 복사서비스하고 있습니다. 위의 원문복사신청 또는 장바구니 담기를 통하여 원문복사서비스 이용이 가능합니다.

이 논문과 함께 출판된 논문 + 더보기