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A kind of chicken gene agk methylation detection method and its application

A detection method and methylation technology, which can be applied in the field of genetic engineering and can solve problems such as the methylation regulation that has not yet existed

Active Publication Date: 2019-06-04
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report on the methylation regulation of AGK gene in Salmonella enteritidis infection

Method used

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  • A kind of chicken gene agk methylation detection method and its application
  • A kind of chicken gene agk methylation detection method and its application
  • A kind of chicken gene agk methylation detection method and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0019] A chicken gene AGK methylation detection method, the method used is nested PCR, the chicken AGK gene specific methylation primers used, and the sequence is shown in the following table:

[0020]

[0021] Choose 6-10 Shouguang chickens negative for Salmonella enteritidis, and the test group will be inoculated with 10 bacteria. 9 Cfu of Salmonella enteritidis bacteria solution 0.3mL, the control group was inoculated with 0.3mL PBS buffer solution, cecal samples were collected 14 days after the inoculation, and genomic DNA was extracted. The test group and the control group each took 3-5.

[0022] The chicken genomic DNA was modified with sulfite with QIAGEN’s methylation modification kit to convert the unmethylated cytosine position into uracil, and the methylation site was due to 5′-methylcytosine Protection without change.

[0023] The first pair of the above primers is the first pair of primers, and the second pair is the second pair of primers. The above-mentioned two pairs ...

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Abstract

The invention relates to the technical field of genetic engineering and provides a chicken gene AGK methylation detection method and the application thereof. According to the method, a specific primer for chicken gene AGK methylation detection and nest-PCR conductions matched with the primer are provided. By means of the primer and the method, methylation of AGK genes of chicken infected with salmonella enteritidis can be detected accurately, and theoretical basis can be provided for chicken salmonella enteritidis resistance with the detection result as the molecular marker.

Description

Technical field [0001] The invention relates to the technical field of genetic engineering, and provides a chicken gene AGK methylation detection method and its application. Background technique [0002] Chicken Salmonella enteritidis is a common disease in poultry production. Salmonella enteritidis can enter egg products through eggshells and can be deposited in the reproductive tract for reproduction. It has been previously reported that most food poisoning incidents in developed countries such as the Americas are caused by Salmonella in poultry, and among them, the main pathogen is Salmonella enteritidis. Salmonella enteritidis has a major impact on poultry production, causing disease in livestock and poultry, and is a food-borne pathogen. It can pose a great threat to human health through poultry by-products and is one of the most frequently reported pathogenic microorganisms. [0003] The AGK gene plays an important regulatory role in diseases, especially cancer. Studies have...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/10C12R1/42
CPCC12Q1/6888C12Q2600/154Y02A50/30
Inventor 李显耀王莎莎吴桂贤王巧巧
Owner SHANDONG AGRICULTURAL UNIVERSITY