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PAPN gene site-directed modified pig

A technology of site-directed modification and gene, applied in genetic engineering, plant genetic improvement, cells modified by introducing foreign genetic material, etc., can solve the problem of gene regulation network that has not been reported

Inactive Publication Date: 2016-05-04
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the important role of the pAPN gene, its crystal structure has also been resolved (Chenetal2012), but the gene regulatory network of its role has not been reported

Method used

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  • PAPN gene site-directed modified pig
  • PAPN gene site-directed modified pig
  • PAPN gene site-directed modified pig

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Experimental program
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Embodiment Construction

[0040] A method for establishing a pAPN gene site-directed modification pig that can be used for anti-diarrheal disease and for human disease models, comprising the following steps:

[0041] (1) Cloning and sequence analysis of pAPN gene

[0042] Using the genome (GenBankaccessnumber: CT737411.6) and cDNA (GenBankaccessnumber: HQ824547.1) sequence information of pAPN, design primers for cloning, use high-fidelity DNA polymerase to amplify the target fragment, clone it into an expression vector, and sequence it. Genome sequence and cDNA sequence are analyzed to detect their mutation sites;

[0043] (2) Construction and activity analysis of pAPN gene editing vector and construction of Donor vector

[0044] ① Construction of pAPN gene editing vector: Using pAPN genome sequence information and mutation site analysis data, select its first exon as the target sequence, design 2 pairs of TALE protein recognition sequences, and clone them into pTALEN-01 vector;

[0045] ②Constructio...

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Abstract

The invention discloses a pAPN gene site-directed modified pig. The pAPN gene site-directed modified pig is obtained by a plurality of technologies, such as constructing a gene vector of porcine aminopeptidase N (pAPN) by adopting a method in genetic engineering and performing cell transfection. The pAPN gene site-directed modified pig is mainly applied in two aspects: 1, the infection of viral diarrhea and K88 of the pig is thoroughly eliminated from a genetic angle, the disease control cost of pig farming is reduced, the pollution to environment caused by pig farming is reduced, and a method for performing healthy cultivation and reducing the abuse of antibiotics is provided; 2, the gene site-directed modified pig is the trial test before researching the screening and clinic of the pathogenesis and related intervention and treatment of related diseases, such as human cancers.

Description

technical field [0001] The invention relates to the technical field of animal genetic engineering, in particular to a pAPN gene site-directed modification pig. Background technique [0002] 1. The loss caused by piglet diarrhea is huge [0003] Piglet diarrhea is a common and typical multifactorial disease under intensive pig production conditions. In recent years, the morbidity and mortality of piglet diarrhea in my country have remained high, and the mortality rate of many pig farms is almost 100%, which has caused great economic losses. According to incomplete statistics, in 2014, there were about 45 million sows in stock in my country, and nearly 800 million piglets were produced annually, but only about 80% of the pigs were finally listed, and the mortality rate of pigs at different stages reached 20%, and about half of them were due to diarrheal disease. Therefore, piglet diarrhea is extremely harmful to the pig industry. [0004] 2. The pathogen of piglet diarrhea ...

Claims

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Application Information

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IPC IPC(8): C12N15/57C12N15/85C12N5/10C12N15/877A01K67/027
CPCC12N9/485A01K67/027A01K2217/075A01K2227/108A01K2267/03A01K2267/0337A01K2267/0381A01K2267/0387C12N15/8509C12N15/877C12N2015/8581C12N2800/107C12Y304/11002
Inventor 陈建文张运海李运生潘开源韩春杨
Owner ANHUI AGRICULTURAL UNIVERSITY
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