Trichoderma harzianum expansin and coding gene and application thereof
A technology for expanding proteins and genes, applied in the field of molecular biology, can solve the problems of no cellulase activity, difficult detection of activity purification, etc., and achieve obvious synergistic effects and improve degradation efficiency
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[0023] Material preparation:
[0024] The Trichoderma harzianum described in the examples, the strain number is CGMCC No.3.5364, was purchased from China General Microorganism Culture Collection and Management Center.
[0025] 1. Mycelia Preparation
[0026] Trichoderma harzianum spore suspension (1.00±0.05)×10 7 cells / ml inoculated on coated fiber plates, cultured for 36 hours, collected mycelia in several 1.5ml centrifuge tubes, and stored in liquid nitrogen quick-frozen.
[0027] The composition of the fiber plate is as follows: 55wt% powder (the mass ratio of bran in the powder: grass powder is 1:1), 1.5wt% agar, and the balance of water.
[0028] 2. Extraction of total RNA and separation and purification of mRNA
[0029] Weigh 0.2 g of mycelium in a pre-cooled mortar and grind it to powder with liquid nitrogen. The RNA extraction method and the mRNA separation and purification method were operated according to the corresponding instructions about RNeasy Mini Kit and O...
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