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

Inactive Publication Date: 2013-05-29
吴珍芳
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  • Abstract
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  • Application Information

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Problems solved by technology

Although some studies have proved that adding xylanase to feed can improve pigs’ digestion and absorption efficiency of feed nutrients, improve pig growth performance, and reduce pollutant emissions in manure, but adding xylanase to feed increases Feed costs, reducing the profit of pig farming
In addition, since some processes in the feed preparation process usually need to be completed under special conditions (such as high temperature), it is easy to cause some enzyme activity loss of the added xylanase, resulting in unstable or ineffective hydrolysis of xylan in the feed. ideal

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  • Xylanase recombinant expression vector and cultivation method of transgenic pig capable of secreting xylanase
  • Xylanase recombinant expression vector and cultivation method of transgenic pig capable of secreting xylanase
  • Xylanase recombinant expression vector and cultivation method of transgenic pig capable of secreting xylanase

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Embodiment Construction

[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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Abstract

The invention discloses a xylanase recombinant expression vector and a cultivation method of a transgenic pig capable of secreting xylanase. The recombinant expression vector which is regulated and controlled to express by a regulatory element is constructed by steps of splicing external xylanase genes and the regulatory element and inserting the spliced one to a eukaryotic expression vector. According to the invention, external xylanase genes are guided to pig genome successfully through a genetic engineering technology and expressed under control of the regulatory element, so that the pig can secrete xylanase in the digestive tract to eliminate the antinutritional effect of xylan in feed. Compared with a method that xylanase is added to pig feed to eliminate antinutritional effect of xylan, the invention has the advantages that the degrading efficiency of xylanase in pig feed remarkably improved, the digestion and absorption efficiencies to the nutritional matters in the feed are improved, the growing performance of pig is improved, the discharge of pollutants in excrement is reduced, the feed cost of the pig is remarkably lowered, and the pig raising benefit is improved.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to transgenic engineering technology, in particular to a xylanase recombinant expression vector and a method for cultivating transgenic pigs capable of self-secreting xylanase by using the recombinant expression vector. Background technique [0002] The most important ingredients in pig feed are corn and soybean meal, etc. These grain seeds are plant feed materials. Plants contain large amounts of non-starch polysaccharides such as cellulose, hemicellulose, and pectin glycans because they are the main components of plant cell walls. Xylan is the most abundant hemicellulose in plant cell walls, accounting for about 35% of the total dry weight of plant cells, and is the main non-starch polysaccharide in plant feed materials. [0003] Studies have found that pigs cannot produce xylanase on their own. Since the types and quantities of microorganisms parasitic in the digest...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/877A61D19/04A01K67/027
Inventor 刘德武李紫聪蔡更元张茂石俊松周荣吴珍芳
Owner 吴珍芳
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