Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment

B. W. Hazell; V. S. J. Te'o; J. R. Bradner; P. L. Bergquist; K. M. H. Nevalainen

doi:10.1046/j.1472-765x.2000.00715.x

Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment

B. W. Hazell, V. S. J. Te'o, J. R. Bradner, P. L. Bergquist, K. M. H. Nevalainen

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Abstract

Intact conidia of three industrially relevant strains of Trichoderma reesei were effectively transformed by particle bombardment. Transformations were carried out individually with plasmids carrying either the fungal amdS or bacterial hph gene as a selectable marker and by cotransformation with both plasmids. Transformant yields with single plasmids were up to 11 stable transformants per μg DNA at the bombardment distance of 6 cm. Mitotic stability of the transformants was 75-100% and the cotransformation efficiency averaged 92% when the first selection was performed on hygromycin B plates. The entire procedure could be completed in 1 week with the hph marker.

Original language	English
Pages (from-to)	282-286
Number of pages	5
Journal	Letters in Applied Microbiology
Volume	30
Issue number	4
DOIs	https://doi.org/10.1046/j.1472-765x.2000.00715.x
Publication status	Published - 2000

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title = "Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment",

abstract = "Intact conidia of three industrially relevant strains of Trichoderma reesei were effectively transformed by particle bombardment. Transformations were carried out individually with plasmids carrying either the fungal amdS or bacterial hph gene as a selectable marker and by cotransformation with both plasmids. Transformant yields with single plasmids were up to 11 stable transformants per μg DNA at the bombardment distance of 6 cm. Mitotic stability of the transformants was 75-100% and the cotransformation efficiency averaged 92% when the first selection was performed on hygromycin B plates. The entire procedure could be completed in 1 week with the hph marker.",

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AU - Hazell, B. W.

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AU - Bradner, J. R.

AU - Bergquist, P. L.

AU - Nevalainen, K. M. H.

PY - 2000

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N2 - Intact conidia of three industrially relevant strains of Trichoderma reesei were effectively transformed by particle bombardment. Transformations were carried out individually with plasmids carrying either the fungal amdS or bacterial hph gene as a selectable marker and by cotransformation with both plasmids. Transformant yields with single plasmids were up to 11 stable transformants per μg DNA at the bombardment distance of 6 cm. Mitotic stability of the transformants was 75-100% and the cotransformation efficiency averaged 92% when the first selection was performed on hygromycin B plates. The entire procedure could be completed in 1 week with the hph marker.

AB - Intact conidia of three industrially relevant strains of Trichoderma reesei were effectively transformed by particle bombardment. Transformations were carried out individually with plasmids carrying either the fungal amdS or bacterial hph gene as a selectable marker and by cotransformation with both plasmids. Transformant yields with single plasmids were up to 11 stable transformants per μg DNA at the bombardment distance of 6 cm. Mitotic stability of the transformants was 75-100% and the cotransformation efficiency averaged 92% when the first selection was performed on hygromycin B plates. The entire procedure could be completed in 1 week with the hph marker.

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Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment

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