한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제24권5호
  • /
  • Pages.553-559
  • /
  • 1996
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  • 1598-642X(pISSN)
  • /
  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

PelB Signal Sequence로 유도된 재조합 인간 상피세포 증식인자 분비 발현 벡터의 제조

  • 박세철 ((주)유한양행 중앙연구소 생물제품연구실) ;
  • 남정현 ((주)유한양행 중앙연구소 생물제품연구실) ;
  • 김정근 ;
  • 권태종 ((주)유한양행 중앙연구소 생물제품연구실) ;
  • 고인영 ;
  • 유광현 (건국대학교 미생물학과)
  • Park, Se-Cheol ;
  • Nam, Jung-Hyun ;
  • Kim, Jeong-Keun ;
  • Kwon, Tae-Jong ;
  • Ko, In-Young ;
  • You, Kwang-Hyun
  • 발행 : 1996.10.01
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초록

We have designed nucleotide sequences of hEGF structural gene to eliminate the N-terminal methionine residue incorporated during the translation initiation step, and constructed recombinant human epidermal growth factor (rhEGF) secretion plasmids pYHB101, and pYHB2 in which pelB signal sequence-hEGF gene was expressed under the control of the T7, and tac promoter, respectively. We also constructed pYHB1 vector which contains rhEGF gene controlled by T7 promoter. The transformant with pYHB101 showed relatively slow growth pattern compared to the transformant with pYHB1. However, we observed that the transformant with pYHB101 secreted rhEGF of 13 mg/l significantly after 5 hr induction with 1 mM IPTG and that the T7 promoter was more effective than tac promoter when connected to pelB signal sequence. The amount of rhEGF was 14 mg/l under the sub-optimized condition.

키워드

참고문헌

  1. J. Cell. Physiol. v.88 Human epidermal growth factor and proliferlation of human fibroblast. Carpenter G;S Cohen.
  2. J. Biol. Chem. v.263 Cloning of authentic human epidermal growth factor as a bacterial secretion protein and its initial structure-function analysis by site-directed mutageneis. David A.E;R.Matsunami;S.R.Campion;C.D.Stringer;A.Stevens;S.K.Niyogi.
  3. Proc.Natl. Acad. Sci. v.86 Use of Bacillus brevis for efficient synthesis and secretion of human epidermal growth factor. Hideo Y;N.Kazuo;S.Yutaka;K.Atsushi
  4. Proc.Natl. Acad. Sci. v.84 Extent of N-terminal methionine excision from Escherichia coli proteins is governed by the side-chain length of the penultimate amino acid. Hirel P.H;J.M Schmitter;P.Dessen;G.Fayat;S.Blanquet.
  5. Nucleic Acids Res. v.10 Chemical synthesis and cloning of a gene for human β-urogastrone. John S;E.Cook;I.Fotheringham;S.Pheby;R.Derbyshire;M.Eaton;M.Doel;D.Lilley;J.Pardon;T.Patel;H.Lewis;L.Bell.
  6. Science v.254 The N-end rule in bacteria. John W.T;T.Shrader;G.Rocap;A.Varshavsky
  7. Gene. v.45 Directed expression of urogastrone gene in Escherichia coli. Kishimoto F;H.Gomi;M.Kanaoka;T.Nakatani;A.Ito;T.Katoh;H.Agui;S.Sumida;S.Ogino.
  8. J. Biochem. v.97 Secretion of Human Interferon-a induced by using secretion vectors containg a promoter and signal sequence of alkaline phoaphatase gene of Escherichia coli. Miyake T;T.Oka;T.Nishizawa.
  9. Agri. Biol. Chem. v.51 Efficacies of different secretion vectors for secretion of Human Epidermal Growth Factor by Escherichia coli. Oka T;S.Sumi;T.Fuwa;K.Yoda;M.Yamasaki;G.Tamura;T.Miyake.
  10. Bio/technology. v.10 DnaK-mediated alterations in human growth hormone proteins inclusion bodies. Blum P;M.Velligan;N.Lin;A.Matin.
  11. Molecular Cloning, 2nd ed. Sambrook J;E.F.Fritsch;T.Maniatis.
  12. Methods Enzymol v.185 Use of T7 RNA polymerase to direct expression of cloned genes. Studier F.W;A.H.Rosenberg;J.J.Dunn;J.W.Dubendorff.
  13. Proc.Natl. Acad. Sci. v.82 Synthesis and secretion of human epidermal growth factor by Escherichia coli. Takanori O;S.Sakamoto;K.Miyoshi;T.Fuwa;K.Yoda;M.Yamasaki;G.Tamura;T.Miyake
  14. J. Microbiol. Methods. v.21 Construction of a transducible cassette encoding an inducible bacteriophage T7 RNA polymerase gene 1 in Escherichia coli. Tedin K;U.Altanerova;U.Blasi.
  15. Proc.Natl. Acad. Sci. v.80 Chemical synthesis of a gene for human epidermal growth factor urogastrone and its expression in yeast. Urdea M.S;J.P.Merryweather;G.T.Mullenbach;D.Coit;U.Heberlein;P.Valenzucla;P.J.Barr.
  16. J. Bacteriology v.169 Characterization of the Erwinga carotovora pelB gene and its product prctate lyase. Lei S.P;H.C.Lin;S.S.Wang;J.Callaway;G.Wilcox.
  17. TiBtech. v.12 Heterogous protein secretion and the versatile Escherichia coli haemolysin translocator. Blight M.A;I.B.Holland.
  18. App. Microbial. Biotech. v.36 Maximizing the expression of a recombinant gene in Escherichia coli by manipulation of induction time using lactose as inducer. Neubauer P;K.Hofman;O.Holst;B.Mattasson;P.Kruschke.