Xylanase recombinant expression vector and cultivation method of transgenic pig capable of secreting xylanase
A technology of xylanase and expression vector, which is applied in the field of genetic engineering, can solve the problems of loss of enzyme activity, reduction of pig raising profit, unsatisfactory problems, etc., to improve digestion and absorption efficiency, increase pig raising profit, and reduce feed cost Effect
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[0038] The present invention will be further described below in conjunction with specific examples, but the specific examples do not limit the present invention in any way.
[0039] The specific embodiment uses the Aspergillus niger xylanase gene XynB as the exogenous xylanase gene.
[0040] 1. Cloning of Aspergillus niger xylanase gene XynB
[0041] Refer to the Aspergillus niger xylanase gene xynB sequence published in Genbank (Genbank accession number: DQ174549) to design primers xynBF and xynBR to amplify the mature peptide coding sequence.
[0042] The nucleotide sequence of xynBF is shown in SEQ ID NO:1, and the nucleotide sequence of xynBR is shown in SEQ ID NO:2.
[0043] Extract the total RNA of Aspergillus niger and use it as a template for RT-PCR amplification. The RT-PCR reaction program: 50°C for 30 min, 94°C for 2 min; 94°C for 30 s, 52°C for 30 s, 72°C for 1 min, 35 cycles , and finally extended at 72°C for 10 min.
[0044]RT-PCR products were detected by 1...
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