Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 51, Issue 3, Pages 393-399Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0167-7012(02)00126-4
Keywords
biolistic transformation; Hepta adaptor; Trichoderma reesei
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Effective biolistic transformation of intact conidia from the filamentous fungus Trichoderma reesei was achieved using the Bio-Rad Hepta Adaptor system with seven barrels for particle launch. Transformation frequencies of up to 39 colonies per mug of circular DNA and 37 colonies per mug of linear DNA were obtained at an optimal target distance of 3 cm and a helium pressure of 1350 psi. These values are about 3.5- to 6-fold higher than transformant yields reported earlier for T reesei using the hygromycin phosphotransferase (hph) gene conferring resistance to the antibiotic hygromycin B as a selectable marker in combination with the PDS-1000/He single barrel system. High mitotic stability of the transformants (98-100%) was demonstrated. The Hepta Adaptor device allowing bombardment of seven lots of conidia in a single plate offers clear advantage in terms of transformant numbers over the single barrel system where target cells are restricted to the center of the plate. (C) 2002 Elsevier Science B.V. All rights reserved.
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