Biolistic transformation of Trichoderma reesei using the Bio-Rad seven barrels Hepta Adaptor system.
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 microg of circular DNA and 37 colonies per microg 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.
Te'o
V S J
VS
Department of Biological Sciences, Macquarie University, Sydney, NSW 2109, Australia. jteo@rna.bio.mq.edu.au
Bergquist
P L
PL
Nevalainen
K M H
KM
D002480:Cellulase / Q000235:genetics; D043366:Cellulose 1,4-beta-Cellobiosidase; D004270:DNA, Circular / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D016681:Genome, Fungal; D009172:Mycology / Q000379:methods; D010957:Plasmids / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D014170:Transformation, Genetic; D014242:Trichoderma / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development
D004270:DNA, Circular; D004271:DNA, Fungal; D011933:Reagent Kits, Diagnostic; D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase