Encoding gene of glycoside hydrolase for rapid hydrolysizing xylan to generate single xylose, and application of encoding gene
A technology of glycoside hydrolase and xylan, applied in the field of coding genes of glycoside hydrolase
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[0057] This example is used to illustrate the construction of the recombinant strain containing the Ttxy43 gene of the present invention.
[0058] (1) Acquisition of Ttxy43 gene
[0059] The nucleic acid sequence shown in SEQ ID NO:2 was synthesized.
[0060] Design of specific primers Ttxy43-F / Ttxy43-R
[0061] Ttxy43-F: CGGAATTCATGGCTCCATTGATCACCAA (SEQ ID NO: 8)
[0062] Ttxy43-R: GGGGTACCTTACTCTGGCTTCTCGGTA (SEQ ID NO: 9)
[0063] Using the nucleic acid sequence shown in SEQ ID NO: 2 as a template and Ttxy43-F / Ttxy43-R as primers for PCR amplification, the reaction parameters of PCR are: denaturation at 95°C for 5 minutes, denaturation at 95°C for 30 sec, annealing at 60°C for 30 sec, Extended at 72°C, and continued to extend at 72°C for 10 min after 30 cycles to obtain a Ttxy43 gene fragment (SEQ ID NO: 2).
[0064] (2) Construction of recombinant strains
[0065]Use restriction endonucleases EcoR I and Kpn I to digest the Ttxy43 gene fragment and the pPICZαA plasmid...
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