Pig intestinal epithelial cell promoter SIECP and application thereof

A promoter and molecular technology, applied in the biological field, can solve problems such as the cause of disease and effective treatment methods that need to be further explored, and achieve the effect of wide application prospects

Inactive Publication Date: 2019-12-27
CHINA AGRI UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

For example, the incidence of ulcerative colitis in my country is about 1 in 10,000, and viral enteritis affects 51 million people in the United States every year, but the specific causes and effective treatment methods need to be further explored

Method used

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  • Pig intestinal epithelial cell promoter SIECP and application thereof
  • Pig intestinal epithelial cell promoter SIECP and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] This example provides a DNA fragment derived from Chinese experimental minipigs, which has promoter activity and is denoted as SIECP. For the detection flow chart, see figure 1 .

[0057] 1. Acquisition of target fragments

[0058] Using the Chinese experimental minipig genome as a template, PCR amplification was performed to obtain PCR products. The primers used are as follows:

[0059] The upstream primer is 5'-CGacgcgtGTTTGCTTGGCTTGATCAGCAG-3' (containing MluI recognition sequence);

[0060] The downstream primer is 5'-CTAgctagcATTGACCAGCTCCCCTCTAGTG-3' (containing NheI recognition sequence).

[0061] 2. Construction of pGL3-SIECP recombinant plasmid

[0062] The PCR product obtained in step 1 was double-digested with MluI and NheI, and the resulting fragment was connected to the large vector fragment obtained by double-digestion of the pGL3-Basic plasmid with MluI and NheI to obtain a recombinant plasmid, and the recombinant plasmid with the correct sequence was ...

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Abstract

The invention discloses a pig intestinal epithelial cell promoter SIECP and application thereof. the disclosed pig intestinal epithelial cell promoter SIECP is a DNA molecule shown in a sequence 1 ina sequence table. The SIECP can activate the target gene expression in intestinal epithelial cells, and can be used for cultivating transgenic animals, theoretical basis is provided for constructing stable and efficient intestine transcription regulatory elements, reference value is achieved for the production of environment-friendly transgenic animals and the gene therapy research of intestine diseases, and wide application prospects in the field of life science and intestine disease treatment are achieved.

Description

technical field [0001] The invention relates to a promoter SIECP of pig small intestinal epithelial cells and its application in the field of biotechnology. Background technique [0002] 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 intestinal cancer and other diseases. For example, the incidence of ulcerative colitis in my country is about 1 in 10,000, and 51 million people are infected with viral enteritis every year in the United States, but the specific causes and effective treatment methods need to be further explored. Therefore, it is of great significance to ascertain the pathogenesis of various types of enteritis and find suitable model animals. [0003] Efficient and stable expression of exogenous genes in target tissues is an important prerequisite for transgenic animal research and gene therapy. The expression of exogenous genes such as pathogen receptors ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N5/10C12N15/11C12N15/85
CPCC07K14/47C12N15/85
Inventor 张然白兰
Owner CHINA AGRI UNIV
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