Mannanases and recombinant expression bacterial strain thereof
A technology of mannanase and recombinant vector, which is applied in the field of microbial engineering and can solve problems such as intolerance to pepsin
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[0026] The method of the present invention will be further described below in conjunction with examples. Experimental methods that do not specify specific conditions in the examples can usually be based on conventional conditions, as described in the "Molecular Cloning Experiment Guide" written by J. Sambrook (Sambrook) etc. operating conditions, or as recommended by the manufacturer. Those skilled in the art can better understand and master the present invention with the help of the examples. However, the scope of protection and claims of the present invention is not limited to the examples provided.
[0027] 1. Cloning of Penicillium decumbens mannanase gene
[0028] 1.1 Extraction of total DNA
[0029] Culture Penicillium decumbens (P. decumberns) overnight, put an appropriate amount of bacteria into a centrifuge tube, centrifuge at 13,000 rpm for 5 min, discard the supernatant; add 400 μl of extraction buffer (100 mM Tris-HCl, 100 mM EDTA, 250 mM NaCl, 1%SDS); then add ...
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