Preparation method for recombination strains for generating Beta glucosaccharase
A technology of glucosidase and construction method, which is applied in the field of construction of recombinant bacteria producing β-glucosidase, which can solve the problems of low enzyme activity, difficult extraction, complex components of β-glucosidase, etc.
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[0018] 1. Extraction of Aspergillus niger mRNA
[0019] 1. Aspergillus niger total RNA extraction:
[0020] (1) After grinding the mycelium into powder in liquid nitrogen, add 1 mL of Trizol liquid nitrogen per 50-100 mg of mycelium for grinding, and note that the total sample volume cannot exceed 10% of the volume of Trizol used.
[0021] (2) The grinding solution was placed at room temperature for 5 minutes, then chloroform was added at a ratio of 0.2 mL per 1 mL of Trizol solution, the centrifuge tube was tightly capped, and the centrifuge tube was shaken vigorously by hand for 15 seconds.
[0022] (3) Take the upper aqueous phase into a new centrifuge tube, add isopropanol at a ratio of 0.5 mL of isopropanol per mL of Trizol solution, place at room temperature for 10 minutes, and centrifuge at 12,000g for 10 minutes.
[0023] (4) Discard the supernatant, add at least 1 mL of 75% ethanol per 1 mL of Trizol solution, mix by vortexing, and centrifuge at 7500g for 5 minutes a...
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