Short Communication

Current Genetics

, Volume 25, Issue 6, pp 567-570

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

  • Robert L. MachAffiliated withAbteilung für Mikrobielle Biochemie, Institut für Biochemische Technologie und Mikrobiologie, TU Wien
  • , Martin SchindlerAffiliated withAbteilung für Mikrobielle Biochemie, Institut für Biochemische Technologie und Mikrobiologie, TU Wien
  • , Christian P. KubicekAffiliated withAbteilung für Mikrobielle Biochemie, Institut für Biochemische Technologie und Mikrobiologie, TU Wien

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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.

Key words

Trichoderma reesei Transformation Hygromycin B Hygromycin B phosphotransferase Pyruvate Kinase Promoter Terminator Dominant selection marker