Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Development of a Multicopy Integration Vector in Yarrowia lipolytica

Yarrowia lipolytica의 Multicopy Integration Vector 개발

  • 김정윤 (충남대학교 자연과학대학 미생물학과) ;
  • 우문희 (충남대학교 자연과학대학 미생물학과) ;
  • Published : 1995.10.01
Download PDF
⟨ Previous Next ⟩

Abstract

Multicopy integration vector is a very useful vector system in that they can be integrated into chromosomal DNA in several copies and stably maintained under non-selective conditions. To develop a multicopy integration vector system in the yeast Yarrowia lipolytica, P-type ribosomal DNA was cloned from Y lipolytica. A HindIII-BglII fragment of the cloned rDNA and a promoterless URA3 gene were inserted into pGEM1, generating multicopy integration vectors, pMIYL-1 and pMIYL-2. The rDNA fragment is for targeted homologous recombination between the vector and the chromosomal DNA of Y. lipolytica, and the promoterless URA3 gene is a defective selection marker for inducing multicopy integration. pMIYL-1 and pMIYL-2 have an unique restriction enzyme site, KpnI, and two unique restriction enzyme sites, KpnI and EcoRI, repectively, which can be used for targeting of the vectors into the rDNA of Y. lipolytica chromosomal DNA. After transformation of the vectors into Y. lipolytica, copy number and stability were analyzed by Southern hybridization. The vectors were found to be present in less than 5 copies per cell and were stably maintained during growth in non-selective media.

Keywords

References

  1. Meth. Enzymol. v.185 Expression of heterologous proteins in Escherichia coli Gold,L.
  2. Meth, Enzymol. v.185 Inducible expression of regulactory genes in Bacillus subtilis Henner,D.J.
  3. Trens Biotechnol. v.10 Bacillus subtilis and its relatives: molecular biological and industrial work horses Harwood,C.R.
  4. Meth. Enzymol. v.10 Heterologous gene expression in yeast. Bitter,G.A.
  5. Yeast Genetic Engineering, Butterworths Secretion of heterologous proteins directed by the yeast α-factor leader Brake,A.J.;P.J.Barr(ed.);P.Valenzuela(ed.)
  6. Trends Biotechnol. v.5 The production of mammalian proteins in Saccharomyces cerevisiae Kingsman,S.M.;A.J.Kingsman;J.Mellor
  7. Recombinant DNA Technology and Applications Expression of heterologous protein in yeast Marino,M.H.;A.Prokop(ed.);R.K.Bajpai(ed.);C.Ho(ed.)
  8. Yeast v.8 Foreign gene expression in yeast: a review Romanos,M.A.;C.A.Scorer;J.J.Clare
  9. Dev. Ind. Microbiol. v.29 Development of the methylotrophic yeast, Pichia pastoris, as a host system for the production of foreign proteins Cregg,J.M.;K.R.Madden
  10. Dev. Ind. Microbiol. v.29 Expression and secretion of bovine prochymosin in Yarrowia lipolytica Franke,A.E.;F.S.Kaczmarek;M.E.Eisehard;K.F.Geoghegan;D.E.Danley;J.R.DeZeeuw;M.M.O'Donnell;M.G.Gollaher;L.S.Davidow
  11. Bio/Technol. v.8 Kluyveromyces as a host for heterologous gene expression : Expression and secretion of prochymosin Van den Berg;J.A.;K.J.van der Laken;A.J.J.van Ooyen;T.C.H.M.Renniers;K.Rietveld;A.Schaap;A.J.Brake;R.J.Bishop;K.Schultz;D.Moyer;M.Richman;J.R.Shuster
  12. Agric. Biol. Chem. v.40 Production and some enzymatic properties of alkaline protease of Candida lipolytica Tobe,S.;T.Takami;S.Ikeda;K.Mitsugi
  13. J. Cell. Biol. v.124 A phosphatidylinositol/phosphatidyl cholin transfer protein is required for differentiation of the dimorphic yeast Yarrowia lipolytica from the yeast to the mycelial form Lopez,M.C.;J.M.Nicaud;H.B.Skinner;C.Vergnolle;J.C.Kader;V.A.Bankaitis;C.Gaillardin
  14. U.S.A. Patent No. 4407953 Fermentation process for production of alpha-isopropylmalic acid DeZeeuw,J.R.;I.Stasko
  15. U,S,A, Patent No. 3736229 Fermentation process for the production of erythritol DeZeeuw,J.R.;E.J.Tynan
  16. U.S.A. Patent No. 3756917 Fermentation process for the production of eryfhritol DeZeeuw,J.R.;E.J.Tynan
  17. Enz. Microbiol Technol. v.4 Preliminary genetic studies on a citric acid producing strain of Saccharomycopsis lipolytica Shah,D.N.;A.P.Purohit;K.S.Sriprakash
  18. Z. Allg. Mikrobiol. v.24 Improved conditions for mating of the yeast Saccharomycopsis lipolytica Barth,J.B.;R.K.Mortimer
  19. Curr. Genet. v.5 Genetic and biotechemical studies of N-alkane non-utilizing mutants of Saccharomycopsis lipolytica Bassel,J.B.;R.K.Mortimer
  20. Mol. Gen. Genet. v.188 Development of the genetic map of the yeast Saccharomycopsis lipolytica Ogrydziak,D.;J.Bassel;R.Mortimer
  21. J. Bacteriol. v.169 Processing and secretion of the Yarrowia lipolytica RNase. Cheng,S.K.;D.M.ogrydziak
  22. J. Biotechnol. v.19 Use of ars18 based vectors to increase protein production in Yarrowia lipolytica Nicaud,J.M.;P.Fournier,C.;LaBonnardiere,M.Chasles;C.Gaillardin
  23. Proc. Natl. Acad. Sci. v.90 Colocalization of centromeric and replicative functions on autonomously replication sequences isolated from the yeast Yarrowia lipolytica. Fournier,P.;A.Abbas;B.Kudla;D.M.Ogrydziak;D.Yaver;J.W.Xuan;A.peito;A.M.Ribet;C.Feynerol;F.He;C.Gaillardin
  24. Yeast v.7 Scarcity of ars sequences isolated in a morphogenesis mutant of the yeast Yarrowia lopolytica Fournier,P.;L.Guyaneux;M.Chasles;C.Gaillardin
  25. Mol. Gen. Genet. v.237 Analysis of regions essential for the function of chromosomal replicator sequences from Yarrowia lipolytica Matsuoka,M.;M.Matsubara;H.Daidoh;T.Imanaka;K.Uchida;S.Aiba
  26. Curr. Genet v.21 Multiple-copy integration of the α-galactosidase gene from Cyamopsis tetragonoloba into the ribosomal DNA of Kluyveromyces lactis Bergkamp,R.J.M.;I.M.Kool;R.H.Geerse;R.J.Planta
  27. Gene v.79 High-copy-number integration into the robosomal DNA of Saccharomyces cerevisiae: a new vector for high-level expression Lopez,T.S.;J.Klootwijk;A.E.Veenstra;P.C.van der Aar;H.van Heerikhuizen;H.A.raue;R.J.Palnta
  28. Gene v.105 Mechanism of high copy-number integration into the robosomal DNA of Saccharomyces cerevisiae Lopez,T.S.;F.A.J.Hakkaart;B.L.Koerts;H.A.Raue;R.J.Planta
  29. A Laboratory Manual, 2nd ed. Molecular Cloning Sambrook,J.;E.F.Fritsch;T.Maniatis
  30. Curr. Genet. v.10 Integrative transformation of the yeast Yarrowia lipolytica Davidow,L.S.;D.Apostilakos;M.M.O'Donnell;A.R.Proctor;D.M.Ogrydziak;R.A.Wing;I.Stasko;J.R.Dezeeuw
  31. Proc. Natl. Acad. Sic. USA v.91 The froquency of gene targetinf in yeast depends on the number of target copies Wilson,J.H.;W.Y.Leung;G.Bosco;D.Dieu;J.E.Haber
  32. Gene v.39 Heterogeneity in the robosomal RNA genes of the yeast Yarrowia lipolytica: cloning and analysis of two size classes of repeats Van Herikhuizen,H.;A.Ykema;J.Klootwijk;C.Gaillardin;C.Ballas;P.Fournier