Transformation system for hypocrea jecorina (trichoderma reesei) that favors homologous integration and employs reusable bidirectionally selectable markers

Matthias G. Steiger, Marika Vitikainen, Pekka Uskonen, Kurt Brunner, Gerhard Adam, Tiina Pakula, Merja Penttilä, Markku Saloheimo, Robert L. Mach, Astrid R. Mach-Aigner (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

71 Citations (Scopus)

Abstract

Hypocrea jecorina is an industrially important filamentous fungus due to its effective production of hydrolytic enzymes. It has received increasing interest because of its ability to convert lignocellulosic biomass to monomeric sugars, which can be converted into biofuels or platform chemicals. Genetic engineering of strains is a highly important means of meeting the requirements of tailor-made applications. Therefore, we report the development of a transformation system that allows highly efficient gene targeting by using a tmus53 (human LIG4 homolog) deletion strain. Moreover, it permits the unlimited reuse of the same marker by employing a Cre/loxP-based excision system. Both marker insertion and marker excision can be positively selected for by combining resistance to hygromycin B and loss of sensitivity to fluoroacetamide. Finally, the marker pyr4, also positively selectable for insertion and loss, can be used to remove the cre gene.

Original languageEnglish
Pages (from-to)114-121
JournalApplied and Environmental Microbiology
Volume77
Issue number1
DOIs
Publication statusPublished - 1 Jan 2011
MoE publication typeA1 Journal article-refereed

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Hypocrea
Hygromycin B
Trichoderma reesei
Trichoderma
Genetic Engineering
Gene Targeting
Biofuels
Biomass
Fungi
hygromycin B
gene targeting
Enzymes
biofuels
genetic engineering
Genes
sugars
fungi
gene
biomass
enzymes

Cite this

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title = "Transformation system for hypocrea jecorina (trichoderma reesei) that favors homologous integration and employs reusable bidirectionally selectable markers",
abstract = "Hypocrea jecorina is an industrially important filamentous fungus due to its effective production of hydrolytic enzymes. It has received increasing interest because of its ability to convert lignocellulosic biomass to monomeric sugars, which can be converted into biofuels or platform chemicals. Genetic engineering of strains is a highly important means of meeting the requirements of tailor-made applications. Therefore, we report the development of a transformation system that allows highly efficient gene targeting by using a tmus53 (human LIG4 homolog) deletion strain. Moreover, it permits the unlimited reuse of the same marker by employing a Cre/loxP-based excision system. Both marker insertion and marker excision can be positively selected for by combining resistance to hygromycin B and loss of sensitivity to fluoroacetamide. Finally, the marker pyr4, also positively selectable for insertion and loss, can be used to remove the cre gene.",
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Transformation system for hypocrea jecorina (trichoderma reesei) that favors homologous integration and employs reusable bidirectionally selectable markers. / Steiger, Matthias G.; Vitikainen, Marika; Uskonen, Pekka; Brunner, Kurt; Adam, Gerhard; Pakula, Tiina; Penttilä, Merja; Saloheimo, Markku; Mach, Robert L.; Mach-Aigner, Astrid R. (Corresponding Author).

In: Applied and Environmental Microbiology, Vol. 77, No. 1, 01.01.2011, p. 114-121.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Steiger, Matthias G.

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AU - Brunner, Kurt

AU - Adam, Gerhard

AU - Pakula, Tiina

AU - Penttilä, Merja

AU - Saloheimo, Markku

AU - Mach, Robert L.

AU - Mach-Aigner, Astrid R.

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