The Transformation in Trichoderma viride Protoplasts by restriction enzyme mediated integration (REMI)

摘要

The transformation conditions of the protoplasts from Trichoderma viride mediated by restriction enzyme were studied in this paper. The optimum generation conditions of protoplasts were as followed: 8 mg/ml glucanex was added into the phosphate buffer (pH 6.98), the mycelial that cultured for 24 hr was hydrolyzed for 4 hr at 30°C under 40 r/min shaking speed. The protoplast yield was 4.7×10~7 cfu/mg. The regeneneration rate of protoplast was 14.5% on CM medium contained 0.3 mol/L KCl and 0.3 mol/L Inositol. Transformants were obtained by transfering hygromycin B resistance gene into T. viride by restriction enzyme mediated integration (REMI), The preliminary identification of the transformants indicated that the exogenous gene had been integrated into T. viride genome.