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The pgc promoter expressed by porcine small intestinal epithelial cells and its application

A technology of epithelial cells and pig small intestine, applied in the field of animal genetic engineering

Active Publication Date: 2021-04-06
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • The pgc promoter expressed by porcine small intestinal epithelial cells and its application
  • The pgc promoter expressed by porcine small intestinal epithelial cells and its application
  • The pgc promoter expressed by porcine small intestinal epithelial cells and its application

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Embodiment 1

[0026] The acquisition and application of embodiment 1PGC promoter

[0027] Using genetic engineering technology, look up the PGC gene sequence in the Ensembl database, and predict it through Promoter2.0Prediction Server, and get a reliable PGC promoter region. Primers for the PGC promoter were designed using primer premier 5.0, PCR was performed using the Chinese experimental miniature pig genome as a template, and the amplified fragment was ligated with the pGL3-Basic vector. Through sequencing verification, the recombination of the PGC promoter-Luc expression vector was realized. Then the recombinant plasmid was transfected into IPEC-J2 cells, and the luciferase activity was detected. Studies have found that the PGC promoter can achieve the effect of starting Luc (luciferase), see the schematic diagram figure 1 . Methods as below:

[0028] 1. Prediction of PGC reliable promoter region

[0029] The PGC gene sequence was searched through the Ensembl database, and the 900...

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Abstract

The invention provides a PGC promoter expressed by pig small intestinal epithelial cells and its application. The nucleotide sequence of the PGC promoter is shown in SEQ ID No.1. The present invention predicts the PGC reliable promoter region through an online website; chemical synthesis method Synthesize primers; insert the PGC promoter sequence into the pGL3-Basic vector through the restriction enzyme cutting sites MluⅠ and NheⅠ to obtain the PGC-dual luciferase recombinant plasmid; transfect E. coli for plasmid amplification, extract the plasmid, and verify it by sequencing; Transfected into porcine small intestinal epithelial cells, it was found that the porcine PGC promoter can promote the expression of dual luciferase genes in porcine small intestinal epithelial cells, indicating that the promoter can regulate the expression of downstream genes in porcine small intestinal epithelial cells, which is conducive to the construction of stable The high-efficiency small intestine transcriptional regulatory element has application value in the preparation of transgenic animals and gene therapy research of small intestine diseases.

Description

technical field [0001] The invention relates to the field of animal genetic engineering, in particular to a PGC promoter expressed by pig small intestinal epithelial cells and its application. Background technique [0002] The promoter is an important cis-acting element in the gene, and it is also an important part of the expression vector of genetic engineering. It is generally located dozens of bases upstream of the transcription initiation site. With the development of genetic engineering, it is often necessary to construct an expression vector that can express heterologous proteins at a high level, and the promoter has a great influence on the expression level of foreign genes, and is an important element in genetic engineering. Therefore, choosing an appropriate promoter plays a crucial role in research. [0003] The small intestine is not only the main place for digestion and absorption, but also an important immune barrier, with a high incidence of enteritis and inte...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/11C12N15/85C12Q1/6897
CPCC12N9/6481C12N15/85C12N2800/107C12Q1/6897
Inventor 张然白兰
Owner CHINA AGRI UNIV
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