Cloning of acetyl xylan esterase gene (Auaxe) and preparation of recombinase
A technology of acetyl xylan ester and Aspergillus niger acetyl xylan ester, which is applied in the direction of genetic engineering, plant genetic improvement, recombinant DNA technology, etc., to achieve the effect of large industrial application potential and economic value
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[0020] Cloning of Example 1A.usamii E001Auaxe Mature Peptide cDNA Sequence
[0021] Use A.usamii E001 total RNA as a template and Oligo dT-Adaptor Primer as primers to reverse transcribe the first strand of cDNA; use M13Primer M4 and Auaxe-F as primers for the first round of PCR, and the reaction conditions are: 94°C 2min, 30 cycles (94°C for 30s, 51°C for 30s, 72°C for 1min), 72°C for 10min; use Auaxe-F and Auaxe-R as primers for the second round of PCR, and the reaction conditions are: 94°C for 5min, 30 Cycle (94°C for 30s, 56°C for 30s, 72°C for 1min), 72°C for 10min. The two rounds of PCR products were analyzed by 1% agarose gel electrophoresis, the target band was recovered and ligated with pUCm-T (pUCm-T-Auaxe), transformed into Escherichia coli JM109, and sent to Shanghai Sangon for sequencing.
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