Skip to main content
Log in

Sequence of the cloned pyr4 gene of Trichoderma reesei and its use as a homologous selectable marker for transformation

  • Original Articles
  • Published:
Current Genetics Aims and scope Submit manuscript

Summary

We have cloned and sequenced the Trichoderma reesei pyr4 gene encoding orotidine-5′-monophosphate decarboxylase. Comparison of this sequence with that of the equivalent gene from other filamentous fungi suggests that T. reesei is closely related to Cephalosporium acremonium and Neurospora crassa. The cloned pyr4 gene has been used as a homologous selectable marker for transformation of T. reesei. The majority of transformants obtained with circular plasmid were mitotically unstable and contained non-integrated plasmid molecules, sometimes in addition to plasmid integrated in the genome, Linearization of plasmid prior to transformation decreased the transformation frequency but increased the proportion of stable transformation obtained.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Akins RA, Lambowitz AM (1985) Mol Cell Biol 5: 2272–2278

    Google Scholar 

  • Blallance DJ (1986) Yeast 2: 229–236

    Google Scholar 

  • Ballance DJ, Buxton FP, Turner G (1983) Biophys Res Commun 112 (1): 284–289

    Google Scholar 

  • Beggs JD (1978) Nature 275: 104–109

    Google Scholar 

  • Berka RM, Ward M, Wilson LJ, Hayenga KJ, Kodama KH, Carlomagno LP, Thompson SA (1990) Gene 86: 153–162

    Google Scholar 

  • Boeke JD, LaCroute F, Fink GR (1984) Mol Gen Genet 197: 345–346

    Google Scholar 

  • Buxton FP, Radford A (1983) Mol Gen Genet 190: 403–405

    Google Scholar 

  • Buxton FP, Gwynne DI, Davies RW (1985) Gene 37: 207–214

    Google Scholar 

  • Cantoral JM, Díez B, Barredo JL, Alvarez E, Martín JF (1987) Bio-Technol 5: 494–497

    Google Scholar 

  • Cullen D, Gray GL, Wilson LJ, Hayenga KJ, Lamsa MH, Rey MW, Norton S, Berka RM (1987) Bio-Technol 5: 369–376

    Google Scholar 

  • de Ruiter-Jacobs YMJT, Broekhuijsen M, Unkles SE, Campbell EI, Kingshorn JR, Contreras R, Pouwels PH, Hondel CAMJJ van den (1989) Curr Genet 16: 159–163

    Google Scholar 

  • Gruber F, Visser J, Kubicek P, de Graaff LH (1990) Curr Genet 18: 71–76

    Google Scholar 

  • Gurr S, Unkles S, Kingshorn J (1987) The structure and organization of nuclear genes of filamentous fungi. In: Kinghorn J (ed) Gene structure in eukaryotic microbes. IRL Press, Oxford pp 93–139

    Google Scholar 

  • Mandels M (1985) Biochem Soc Trans 13: 414–416

    Google Scholar 

  • Mandels M, Weber J, Parizek R (1971) Appl Microbiol 21: 152–154

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Montenecourt BS (1983) TIBTECH 1: 156–161

    Google Scholar 

  • Newbury SF, Glazebrook JA, Radford A (1986) Gene 43: 51–58

    Google Scholar 

  • Oakley BR, Rinehart JE, Mitchell BL, Oakley CE, Carmona C, Gray GL, May GS (1987) Gene 61: 385–399

    Google Scholar 

  • Penttilä M, Nevalainen H, Ratto M, Salminen E, Knowles J (1987) Gene 61: 155–164

    Google Scholar 

  • Rowlands RT, Turner G (1973) Mol Gene Genet 126: 201–216

    Google Scholar 

  • Sanger F, Milken S, Coulson AR (1977) Proc Natl Acad Sci USA 74: 5463–5467

    Google Scholar 

  • Sheir-Neiss G, Montenecourt BS (1984) Appl Microbiol Biotechnol 20: 46–53

    Google Scholar 

  • Stohl LL, Lambowitz AM (1983) Proc Natl Acad Sci USA 80: 1058–1062

    Google Scholar 

  • Stohl LL, Akins RA, Lambowitz AM (1984) Nucleic Acids Res 12: 6169–6178

    Google Scholar 

  • Timberlake WE, Barnard EC (1981) Cell 26: 29–37

    Google Scholar 

  • van Heeswijck R (1986) Carlsberg Res Commun 51: 433–443

    Google Scholar 

  • Vian A, Peñalva MA (1989) Nucleic Acids Res 17: 8874

    Google Scholar 

  • Vian A, Peñalva MA (1990) Curr Genet 17: 223–227

    Google Scholar 

  • Vieira J, Messing J (1987) Methods Enzymol 153: 3

    Google Scholar 

  • Vogel HJ (1956) Microbiol Genet Bull 13: 42–43

    Google Scholar 

  • Wilson EM, Franke CA, Black ME, Hruby DE (1989) Gene 77: 69–78

    Google Scholar 

  • Wilson L, Carmona C, Ward M (1988) Nucleic Acids Res 16: 2339

    Google Scholar 

  • Yannisch-Perron C, Vieira J, Messing J (1985) Gene 33: 103–119

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by M.S. Esposito

Rights and permissions

Reprints and permissions

About this article

Cite this article

Smith, J.L., Bayliss, F.T. & Ward, M. Sequence of the cloned pyr4 gene of Trichoderma reesei and its use as a homologous selectable marker for transformation. Curr Genet 19, 27–33 (1991). https://doi.org/10.1007/BF00362084

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00362084

Key words

Navigation