ReviewCurrent genetics2005
Sequential gene deletions in Hypocrea jecorina using a single blaster cassette.
Review in Current genetics, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 62 citations in OpenAlex.
- Conserved storage-carbohydrate metabolic modules are rewired during germination ofFrontiers in fungal biology · 2026Article
- Considerations for Domestication of Novel Strains of Filamentous Fungi.ACS synthetic biology · 2025Review
- Establishment of a selectable marker recycling system for iterative gene editing inSynthetic and systems biotechnology · 2024Article
- URA3 as a Selectable Marker for Disruption and Functional Assessment ofJournal of fungi (Basel, Switzerland) · 2023Article
- Cloning and screening of the putative hexokinase genes from Rhizopus oryzae and their heterologous expression in Saccharomyces cerevisiae.Molecular biology reports · 2022Article
- Development of a powerful synthetic hybrid promoter to improve the cellulase system of Trichoderma reesei for efficient saccharification of corncob residues.Microbial cell factories · 2022Article
- An overview on current molecular tools for heterologous gene expression in Trichoderma.Fungal biology and biotechnology · 2021Review
- A copper-controlled RNA interference system for reversible silencing of target genes inBiotechnology for biofuels · 2018Article
- A versatile system for fast screening and isolation ofBiotechnology for biofuels · 2018Article
- Production of highly efficient cellulase mixtures by genetically exploiting the potentials of Trichoderma reesei endogenous cellulases for hydrolysis of corncob residues.Microbial cell factories · 2017Article
- Omics Analyses of Trichoderma reesei CBS999.97 and QM6a Indicate the Relevance of Female Fertility to Carbohydrate-Active Enzyme and Transporter Levels.Applied and environmental microbiology · 2017Article
- CRISPR/Cas9-mediated efficient genome editing via blastospore-based transformation in entomopathogenic fungus Beauveria bassiana.Scientific reports · 2017Article
- Expression of Talaromyces thermophilus lipase gene in Trichoderma reesei by homologous recombination at the cbh1 locus.Journal of industrial microbiology & biotechnology · 2017Article
- Production of the versatile cellulase for cellulose bioconversion and cellulase inducer synthesis by genetic improvement ofBiotechnology for biofuels · 2017Article
- The N-acetylglucosamine catabolic gene cluster in Trichoderma reesei is controlled by the Ndt80-like transcription factor RON1.Molecular microbiology · 2016Article
- Characterization and Strain Improvement of a Hypercellulytic Variant, Trichoderma reesei SN1, by Genetic Engineering for Optimized Cellulase Production in Biomass Conversion Improvement.Frontiers in microbiology · 2016Article
- Novel Strategies for Genomic Manipulation of Trichoderma reesei with the Purpose of Strain Engineering.Applied and environmental microbiology · 2015Article
- Article
- A constitutive expression system for glycosyl hydrolase family 7 cellobiohydrolases in Hypocrea jecorina.Biotechnology for biofuels · 2015Article
- A novel platform for heterologous gene expression in Trichoderma reesei (Teleomorph Hypocrea jecorina).Microbial cell factories · 2014Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In Hypocrea jecorina (anamorph: Trichoderma reesei) multiple gene deletions are limited by the number of readily available selection markers. We have therefore constructed a blaster cassette which enables successive gene knock-outs in H. jecorina. This 3.5 kb pyr4 blaster cassette contains the H. jecorina pyr4 marker gene encoding orotidine-5'-monophosphate (OMP) decarboxylase flanked by two direct repeats of the Streptoalloteichus hindustanus bleomycin gene (Sh ble), which facilitate the excision of the blaster cassette by homologous recombination after each round of deletion. Functionality of this pyr4 blaster cassette was demonstrated by deletion of the glk1 encoding glucokinase and hxk1 encoding hexokinase. 1.4-1.8 kb of the non-coding flanking regions of both target genes were cloned into the respective blaster cassettes and transformation of a pyr4 negative H. jecorina strain with the two cassettes resulted in 10-13% of the transformants in the deletion of one of the two kinase genes. For excision of the pyr4 blaster cassettes, Deltaglk1 strains were selected for growth in the presence of 5-fluoroorotic acid. Recombination between the two Sh ble elements resulted in uridine auxotrophic strains which retained their respective glucokinase negative phenotype. Subsequent transformation of one of these auxotrophic Deltaglk1 strains with the hexokinase blaster cassette resulted in pyr4 prototrophic strains deleted in both glk1 and hxk1. Deltaglk1 strains showed reduced growth on d-glucose and d-fructose whereas Deltahxkl strains showed reduced compact growth on d-glucose but were unable to grow on d-fructose as carbon source. The double Deltaglk1Deltahxk1 deletion strain was completely unable to grow on either d-glucose or d-fructose.
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