Skip to main content
Log in

Transformation of Trichoderma reesei based on hygromycin B resistance using homologous expression signals

  • Short Communication
  • Published:
Current Genetics Aims and scope Submit manuscript

Abstract

Trichoderma reesei was transformed to hygromycin B resistance using a novel vector, which contains the E. coli hygromycin B phosphotransferase gene (hph) fused between promoter and terminator elements of the homologous Trichoderma pkil (coding for pyruvate kinase) and cbh2 (coding for cellobiohydrolase II) genes, respectively. Transformation frequencies of over 1800–2500 transformants/μg DNA were obtained, which is a 15–20-fold increase over that with pAN7-1, which contains hph between A. nidulans expression signals. Mitotically-stable transformants contained the hph gene and the regulatory sequences of the pkil promoter and the cbh2 terminator integrated into the genome. Evidence for preferentially ectopic integration is given.

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.

References

  • Bailey AM, Mena GL, Herrera-Estrella L (1991) Nucleic Acids Res 19:4273–4278

    Google Scholar 

  • Gonzalez A, Jimenez A, Vasquez D, Davies JE; Schindler D (1978) Biochim Biophys Acta 521:459–469

    Google Scholar 

  • Gritz L, Davies J (1983) Gene 25:179–188

    Google Scholar 

  • Gruber F, Visser J, Kubicek CP, De Graaff L (1990a) Curr Genet 18: 71–76

    Google Scholar 

  • Gruber F, Visser J, Kubicek CP, De Graaff L (1990b) Curr Genet 18:447–451

    Google Scholar 

  • Harkki A, Mäntylä A, Penttilä M, Muttilainen S, Bühler R, Suominen P, Knowles JKC, Nevalainen H (1991) Enzyme Microbiol Technol 13:227–232

    Google Scholar 

  • Herrera-Estrella A, Goldman GH, Van Montagu M (1990) Mol Microbiol 4:839–843

    Google Scholar 

  • Kiuchi N, Naruse a, Yamamoto H, Sekiguchi J (1991) Agric Biol Chem 55:3053–3957

    Google Scholar 

  • Kubicek CP (1992) Adv Biochem Engineer Biotechnol 45:1–28

    Google Scholar 

  • Kubicek CP, Messner R, Gruber F, March RL, Kubicek-Pranz EM (1993) Enzyme Microbiol Technol 15:90–99

    Google Scholar 

  • Kück U, Walz M, Mohr G, Mracek M (1989) Appl. Microbiol Biotechnol 31:358–365

    Google Scholar 

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

    Google Scholar 

  • Punt PJ, Oliver RP, Dingemanse MA; Pouwels PH, Van den Hondel CAMJJ (1987) Gene 56:117–124

    Google Scholar 

  • Rodriquez RJ, Yoder OC (1987) Gene 54:73–81

    Google Scholar 

  • Sambrook J, Frisch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Gold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

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

    Google Scholar 

  • Schindler M, Mach RL, Vollenhofer SK, Hodits R, Gruber F, Visser J, De Graaff L, Kubicek CP (1993) Gene 130:271–275

    Google Scholar 

  • Seitboth B, Gruber F, Messner R, Kubicek CP (1992) J Gen Microbiol 138:1259–1264

    Google Scholar 

  • Singh A, Ursic D, Davies J (1979) Nature 277:146–148

    Google Scholar 

  • Smith JL, Bayliss FT, Ward M (1991) Curr Genet 19:27–33

    Google Scholar 

  • Teeri TT, Lehtovaara P, Kaupinnen S, Salovuori I, Knowles JKC (1987) Gene 51:43–52

    Google Scholar 

  • Ulhao CJ, Vainstain M, Peberdy JF (1992) Curr Genet 21:23–26

    Google Scholar 

  • Unkles S (1992) In: Kinghorn JR, Turner G (eds) Applied molecular genetics of filamentous fungi, pp 29–53

  • Van den Hondel CAMJJ, Punt PJ (1991) In: Peberdy JF, Caten CE, Ogden JE; Bennett JW (eds) Applied molecular genetics of fungi. Cambridge University Press, Cambridge, UK, pp 1–27

    Google Scholar 

  • Wong KKY and Saddler JN (1992) In: Visser J, Kusters-van Someren M, Beldman G, Voragen AGJ (eds) Xylan and Xylanases. Elsevier. Amsterdam New York Tokyo, pp 171–186

    Google Scholar 

  • Yanisch-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 R. J. Schweyen

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mach, R.L., Schindler, M. & Kubicek, C.P. Transformation of Trichoderma reesei based on hygromycin B resistance using homologous expression signals. Curr Genet 25, 567–570 (1994). https://doi.org/10.1007/BF00351679

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00351679

Key words

Navigation