Journal of Microbiology and Biotechnology

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

An Efficient and Stable Method for the Transformation of Heterogeneous Genes into Cephalosporium acremonium Mediated by Agrobacterium tumefaciens

  • XU WEI (Department of Pharmaceutical Engineering, Shenyang Pharmaceutical University) ;
  • ZHU CHUNBAO (Department of Biopharmacevtics, Shanghai Institute of Pharmaceutical Industry(SIPI)) ;
  • ZHU BAOQUAN (Department of Biopharmacevtics, Shanghai Institute of Pharmaceutical Industry(SIPI))
  • Published : 2005.08.01
Download PDF
⟨ Previous Next ⟩

Abstract

A transformation system mediated by Agrobacterium tumefaciens is routinely used for the genetic engineering of plants. Here, we report an efficient and stable method for transformation of heterogeneous genes into an industrial Cephalosporium acremonium by using a similar transformation system established in plants. Both the phleomycin-resistant gene and vgb gene were used as screening markers to confirm the success of transformation by either Southern hybridization or PCR amplification. It was found that acetosyringone (AS) was necessary only for protoplast transformation and the heterogeneous genes transferred were integrated into the genome of C. acremonium. The transformation efficiency obtained with this system was much higher than the conventional techniques used for transformation of C. acremonium.

Keywords

References

  1. Austin, B., R. M. Hall, and B. M. Tyler. 1990. Optimized vectors and selection for transformation of Neurospora crassa and Aspergillus nidulans to bleomycin and phleomycin resistance. Gene 93: 157-162 https://doi.org/10.1016/0378-1119(90)90152-H
  2. Bennett, J. W. and L. L. Lasure. 1991. Growth media, pp. 441-458, In: Bennett, J. W. and L. L. Lasure (eds.). More Gene Manipulations in Fungi. Academic Press, San Diego, CA, U.S.A
  3. Bevan, M. 1984. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 22: 8711-8721
  4. Bundock, P., A. den Dulk-Ras, A. Beijershergen, and P. J. J. Hooykass. 1995. Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae. EMBO J 14: 3206-3214
  5. Groot, M. J., P. Bundock, P. J. Hooykaas, and A. G. Beijersbergen. 1998. Agrobacterium tumefaciens-mediated transformation of filamentous fungi. Nat. Biotechnol. 16: 839-842 https://doi.org/10.1038/nbt0998-839
  6. Hamilton, C. M., A. Frary, C. Lewis, and S. Tanksley. 1996. Stable transfer of intact high molecular weight DNA into plant chromosomes. Proc. Natl. Acad. Sci. USA 93: 9975-9979
  7. Hooykaas, P. J. J. and A. G. M. Beijersbergen. 1994. The virulence system of Agrobacterium tumefaciens. Annu. Rev. Phytopathol. 2: 157-179 https://doi.org/10.1146/annurev.py.02.090164.001105
  8. Isogai, T., M. Yoshida, M. Tanaka, and H. Aoki. 1987. Transformation of Acremonium chrysogenium and Saccharomyces cerevisiae using an antibiotic resistance marker Agric. Biol. Chem. 51: 2321-2329 https://doi.org/10.1271/bbb1961.51.2321
  9. Mattanovish, D., F. Ruker, A. da C. Machado, M. Laimer, F. Regner, H. Steinkellner, G. Himmber, and H. Katinger. 1989. Efficient transformation of Agrobacterium spp. by electroporation. Nucleic Acids Res. 17: 6747 https://doi.org/10.1093/nar/17.16.6747
  10. Piers, K. L., J. D. Heath, X. Liang, K. M. Stephens, and E. W. Nester. 1996. Agrobacterium tumefaciens-mediated transformation of yeast. Proc. Natl. Acad. Sci. USA 93: 1613-1618
  11. Raeder, U. and P. Broda. 1985. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol. 1: 17-20 https://doi.org/10.1111/j.1472-765X.1985.tb01479.x
  12. Zeng, R., C. B. Zhu, W. J. Zhao, and B. Q. Zhu. 1999. Transformation of plasmid p YG715/vgb bearing TrpC promoter in Cephalosporium acremonium. Chinese J Antibiot. 24: 269-272
  13. Sambrook, J., F. Fritsch, and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, U.S.A
  14. Skatrud, P. L., A. J. Teitz, T. D. Ingolia, C. A. Cantwell, D. L. Fisher, J. L. Chapman, and S. W. Queener. 1989. Use of recombinant DNA to improve production of cephalosporin C by C. acremonium. Bio/Technology 7: 477-485 https://doi.org/10.1038/nbt0589-477
  15. Webster, D. A. and C. Y. Liu. 1974. Reduced nicotinamide adenine dinucleotide cytochrome o reductase associated with cytochrome o purified from Vitreoscilla. J. Biol. Chem. 249: 4259-4260
  16. Winans, A. C. 1992. Two way chemical signaling in Agrobacterium-plant interactions. Microbiol. Rev. 56: 12-31