Functional analysis of the cellobiohydrolase I promoter of the filamentous fungus Trichoderma reesei

M. Ilmén, M. L. Onnela, S. Klemsdal, S. Keränen, M. Penttilä

Research output: Contribution to journalArticleScientificpeer-review

85 Citations (Scopus)

Abstract

Functional analysis of the cellulase promoter cbh1 of the filamentous fungus Trichoderma reesei was carried out using the Escherichia coli lacZ gene as a reporter. An assay based on cultivation on solid medium in microtiter plates was developed that allows rapid and reliable semiquantitative analysis of β-galactosidase expression of a large number of transformants. A series of deletions and specifically designed alterations were made covering 2.2kb of the cbh1 promoter. Removal of sequences upstream of nucleotide - 500 in relation to the initiator ATG abolished glucose repression. Mutation of a single hexanucleotide sequence 5'GTGGGG at nucleotide - 720 was sufficient for derepression. This site is similar to the binding sites of the glucose repressors MIG1 of Saccharomyces cerevisiae and CREA/CREI of filamentous fungi. Removal of the glucose repressor site did not affect sophorose induction. Sophorose induction of the promoter was retained even in deletion derivatives lacking sequences upstream of position - 161, which retained about 70 bp upstream of the transcription start point and only 30 bp upstream of the TATA box.

Original languageEnglish
Pages (from-to)303-314
Number of pages12
JournalMolecular and General Genetics
Volume253
Issue number3
DOIs
Publication statusPublished - 1 Dec 1996
MoE publication typeA1 Journal article-refereed

Fingerprint

Cellulose 1,4-beta-Cellobiosidase
Trichoderma
Fungi
Glucose
Galactosidases
TATA Box
Lac Operon
Cellulase
Saccharomyces cerevisiae
Nucleotides
Binding Sites
Escherichia coli
Mutation
sophorose

Keywords

  • Cellulase
  • creA/crel
  • Glucose repression
  • Induction
  • MIG1

Cite this

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title = "Functional analysis of the cellobiohydrolase I promoter of the filamentous fungus Trichoderma reesei",
abstract = "Functional analysis of the cellulase promoter cbh1 of the filamentous fungus Trichoderma reesei was carried out using the Escherichia coli lacZ gene as a reporter. An assay based on cultivation on solid medium in microtiter plates was developed that allows rapid and reliable semiquantitative analysis of β-galactosidase expression of a large number of transformants. A series of deletions and specifically designed alterations were made covering 2.2kb of the cbh1 promoter. Removal of sequences upstream of nucleotide - 500 in relation to the initiator ATG abolished glucose repression. Mutation of a single hexanucleotide sequence 5'GTGGGG at nucleotide - 720 was sufficient for derepression. This site is similar to the binding sites of the glucose repressors MIG1 of Saccharomyces cerevisiae and CREA/CREI of filamentous fungi. Removal of the glucose repressor site did not affect sophorose induction. Sophorose induction of the promoter was retained even in deletion derivatives lacking sequences upstream of position - 161, which retained about 70 bp upstream of the transcription start point and only 30 bp upstream of the TATA box.",
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Functional analysis of the cellobiohydrolase I promoter of the filamentous fungus Trichoderma reesei. / Ilmén, M.; Onnela, M. L.; Klemsdal, S.; Keränen, S.; Penttilä, M.

In: Molecular and General Genetics, Vol. 253, No. 3, 01.12.1996, p. 303-314.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Ilmén, M.

AU - Onnela, M. L.

AU - Klemsdal, S.

AU - Keränen, S.

AU - Penttilä, M.

PY - 1996/12/1

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