Biotechnology and Bioprocess Engineering:BBE

  • 제5권1호
  • /
  • Pages.7-12
  • /
  • 2000
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  • 1226-8372(pISSN)
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  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Expression of Mouse $\alpha-Amylase$ Gene in Methylotrophic Yeast Pichia pastoris

  • Uehara Hiroyuki (Lab. of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University) ;
  • Choi Du Bok (Institute of Life Science, Chosun University) ;
  • Park Enoch Y. (Lab. of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University) ;
  • Okabe Mitsuyasu (Lab. of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University)
  • 발행 : 2000.01.01
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초록

The expression of the mouse $\alpha-amylase$ gene in the methylotrophic yeast, P pastoris was investigated. The mouse $\alpha-amylase$ gene was inserted into the multi-cloning site of a Pichi a expression vector, pPIC9, yielding a new expression vector pME624. The plasmid pME624 was digested with SalI or BglII, and was introduced into P. pastoris strain GSl15 by the PEG1000 method. Fifty-three transformants were obtained by the transplacement of pME624 digested with SaiII or BglII into the HIS4locus $(38\;of\;Mut^+\;clone)$ or into the AOX1 locus $(15\;of\;Mut^s\;clone)$. Southern blot was carried out in 11 transformants, which showed that the mouse $\alpha-amylase$ gene was integrated into the Pichia chromosome. When the second screening was performed in shaker culture, transformant G2 showed the highest $\alpha-amylase$ activity, 290 units/ml after 3-day culture, among 53 transformants. When this expression level of the mouse $\alpha-amylase$ gene is compared with that in recombinant Saccharomyces cerevisiae harboring a plasmid encoding the same mouse $\alpha-amylase$ gene, the specific enzyme activity is eight fold higher than that of the recombinant S. cerevisiae.

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참고문헌

  1. Yeast v.8 Foreign gene expression in yeast: a review Romanos, M. A.;C. A. Scorer;J. J. Clare
  2. J. Biol. Chem. v.265 Characterization of recombinant tick anticoagulant peptide Neeper, M. P.;L. Waxman;D. E. Smith;C. A. Schulman;M. Sardana;R. E. Ellis;L. W. Schaffer;P. K. S. Siegal;G. P. Vlasuk
  3. Phchia pastoris. Bio/Technology v.12 High-level secretion and very efficient isotopic labeling of tick anticoagulant peptide (TAP) expressed in the methylotrophic yeast Laroche, Y.;V. Storme;J. D. Meutter;M. Lauwereys
  4. Agri. Biol. Chem. v.44 Oxidation of methanol by the yeast Pichia pastoris. Purification and properties of alcohol oxidase Couderc, R;J. Baratti
  5. Bio/Technology v.11 Recent advances in the expression of foreign genes in Pichia pastoris Cregg, J. M.;S. V. Thomas;C. R. William
  6. Pichia pastoris. Bio/Technology v.5 High-level secretion of glycosylated invertase in the methylotrophic yeast Tschopp, J. F.;G. Sverlow;R. Kosson;W. Craig;L. Grinna
  7. Biotechnol. Bioeng. v.41 Comparison of three different promoter systems for secretory α-amylase production in fed-batch cultures of recombinant Saccharomyces cerevisiae Park, Y.;S. Shiba;S. Iijima;T. Kobayashi
  8. Mol. Cell. Biol. v.5 Pichia pastoris as a host system for transformations Cregg, J. M.;K. J. Barringer;A. Y. Hessler;K. R. Madden
  9. In: C. Gutherie and G. R. Fink, (eds.) Guide to Yeast Genetics and Molecular Biology Recovery of plasmids from yeast into Echerichia coli shuttle vectors Strathern, J. N.;D. R. Higgins
  10. Agric. Biol. Chem. v.51 Construction of an expression and secretion vector for the yeast Saccharomyces cerevisiae Nishizawa, M.;F. Ozawa;F. Hishinuma
  11. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriopahge T4 Laemmli, U. K.
  12. J. Mol. Biol. v.142 Tissue-specific expression of mouse α-amylase genes Schibler, U.;M. Tosi;A. -C. Pittetp;L. Fabiani;P. K. Wellauer
  13. Biotechnol. Bioeng v.44 Improvement of cloned α-amylase gene expression in fed-batch culture of recombinant Saccharomyces cerevisiae by regulating both glucose and ethanol concentrations using a fuzzy controller Shiba, S.;Y. Nishida;Y. Park;S. Iijima;T. Kobayashi
  14. Canadian J. Bot. Supp. v.73 Production of foreign proteins in the yeast Pichia pastoris Cregg, J. M.;D. R. Higgins
  15. Bio/Technology v.9 Yeast systems for the commercial production of heterologous protein Buckholz, R. G.;M. A. G. Gleeson