Development of Trichoderma reesei transformation technology with the aid of an episomal vector system

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

In conventional transformation procedures of filamentous fungi the transformation frequencies have been modest and only integrative vectors have been available. In modern fungal molecular biology, however, screening approaches with the help episomal vectors have advanced the progress of the field greatly. For example, in enzyme discovery or protein engineering it is beneficial to be able to screen for new enzymes or improved mutant enzymes in the ultimate production host of the enzyme. Episomal vector systems have been developed for a few fungal species including the Aspergilli, but Trichoderma reesei has suffered from a lack of such a system. We have developed T. reesei transformation technology for genetic and enzyme screening purposes. The transformation procedure was optimised to improve the transformation frequencies of any vectors. Telomeric episomal vectors were used, and with these we obtained transformation frequencies that are far above those obtained with integrative vectors and are feasible for genetic and enzyme screening purposes. Procedures for plasmid rescue, colony picking and microtiter plate cultivation were also developed. The use of the novel transformation system in enzyme and genetic screening experiments will be discussed.
Original languageEnglish
Title of host publication3rd European Federation of Biotechnology Conference
Subtitle of host publicationPhysiology of Yeasts and Filamentous Fungi PYFF3
Place of PublicationEspoo
PublisherVTT Technical Research Centre of Finland
Pages149
ISBN (Electronic)978-951-38-6314-2
ISBN (Print)978-951-38-6313-5
Publication statusPublished - 2007
Event3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi - Helsinki, Finland
Duration: 13 Jun 200716 Jun 2007

Publication series

NameVTT Symposium
PublisherVTT
Number245
ISSN (Print)0357-9387
ISSN (Electronic)1455-0873

Conference

Conference3rd European Federation of Biotechnology Conference
Abbreviated titlePYFF3
CountryFinland
CityHelsinki
Period13/06/0716/06/07

Fingerprint

Trichoderma reesei
enzymes
screening
protein engineering
Aspergillus
molecular biology
plasmids
mutants
fungi

Cite this

Aro, N., Westerholm-Parvinen, A., Penttilä, M., & Saloheimo, M. (2007). Development of Trichoderma reesei transformation technology with the aid of an episomal vector system. In 3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3 (pp. 149). [P93] Espoo: VTT Technical Research Centre of Finland. VTT Symposium, No. 245
Aro, Nina ; Westerholm-Parvinen, Ann ; Penttilä, Merja ; Saloheimo, Markku. / Development of Trichoderma reesei transformation technology with the aid of an episomal vector system. 3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3. Espoo : VTT Technical Research Centre of Finland, 2007. pp. 149 (VTT Symposium; No. 245).
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Aro, N, Westerholm-Parvinen, A, Penttilä, M & Saloheimo, M 2007, Development of Trichoderma reesei transformation technology with the aid of an episomal vector system. in 3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3., P93, VTT Technical Research Centre of Finland, Espoo, VTT Symposium, no. 245, pp. 149, 3rd European Federation of Biotechnology Conference , Helsinki, Finland, 13/06/07.

Development of Trichoderma reesei transformation technology with the aid of an episomal vector system. / Aro, Nina; Westerholm-Parvinen, Ann; Penttilä, Merja; Saloheimo, Markku.

3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3. Espoo : VTT Technical Research Centre of Finland, 2007. p. 149 P93 (VTT Symposium; No. 245).

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

TY - CHAP

T1 - Development of Trichoderma reesei transformation technology with the aid of an episomal vector system

AU - Aro, Nina

AU - Westerholm-Parvinen, Ann

AU - Penttilä, Merja

AU - Saloheimo, Markku

PY - 2007

Y1 - 2007

N2 - In conventional transformation procedures of filamentous fungi the transformation frequencies have been modest and only integrative vectors have been available. In modern fungal molecular biology, however, screening approaches with the help episomal vectors have advanced the progress of the field greatly. For example, in enzyme discovery or protein engineering it is beneficial to be able to screen for new enzymes or improved mutant enzymes in the ultimate production host of the enzyme. Episomal vector systems have been developed for a few fungal species including the Aspergilli, but Trichoderma reesei has suffered from a lack of such a system. We have developed T. reesei transformation technology for genetic and enzyme screening purposes. The transformation procedure was optimised to improve the transformation frequencies of any vectors. Telomeric episomal vectors were used, and with these we obtained transformation frequencies that are far above those obtained with integrative vectors and are feasible for genetic and enzyme screening purposes. Procedures for plasmid rescue, colony picking and microtiter plate cultivation were also developed. The use of the novel transformation system in enzyme and genetic screening experiments will be discussed.

AB - In conventional transformation procedures of filamentous fungi the transformation frequencies have been modest and only integrative vectors have been available. In modern fungal molecular biology, however, screening approaches with the help episomal vectors have advanced the progress of the field greatly. For example, in enzyme discovery or protein engineering it is beneficial to be able to screen for new enzymes or improved mutant enzymes in the ultimate production host of the enzyme. Episomal vector systems have been developed for a few fungal species including the Aspergilli, but Trichoderma reesei has suffered from a lack of such a system. We have developed T. reesei transformation technology for genetic and enzyme screening purposes. The transformation procedure was optimised to improve the transformation frequencies of any vectors. Telomeric episomal vectors were used, and with these we obtained transformation frequencies that are far above those obtained with integrative vectors and are feasible for genetic and enzyme screening purposes. Procedures for plasmid rescue, colony picking and microtiter plate cultivation were also developed. The use of the novel transformation system in enzyme and genetic screening experiments will be discussed.

M3 - Conference abstract in proceedings

SN - 978-951-38-6313-5

T3 - VTT Symposium

SP - 149

BT - 3rd European Federation of Biotechnology Conference

PB - VTT Technical Research Centre of Finland

CY - Espoo

ER -

Aro N, Westerholm-Parvinen A, Penttilä M, Saloheimo M. Development of Trichoderma reesei transformation technology with the aid of an episomal vector system. In 3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3. Espoo: VTT Technical Research Centre of Finland. 2007. p. 149. P93. (VTT Symposium; No. 245).