SNP marker related to enterotoxigenic Escherichia coli F4 type piglet diarrhea resistance and detection method

A technology for Escherichia coli and piglet diarrhea, applied in the field of molecular biology, can solve the problems of inability to determine the causal variation sites of key genes, difficulty in direct application of piglet diarrhea resistance selection, and no reliable conclusion on the regulation mechanism of Escherichia coli F4 resistance. , to achieve the effect of important economic and social value

Inactive Publication Date: 2018-04-20
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, most studies believe that the receptor gene may be located in the 1Mb interval of chromosome 13 of the pig genome (version 10.2), but the real key gene and its causal mutation site cannot be determined, and the mechanism of E. coli F4 resistance regulation has not yet been reliably determined.
Therefore, it is difficult to directly apply it to piglet diarrhea resistance breeding

Method used

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  • SNP marker related to enterotoxigenic Escherichia coli F4 type piglet diarrhea resistance and detection method
  • SNP marker related to enterotoxigenic Escherichia coli F4 type piglet diarrhea resistance and detection method
  • SNP marker related to enterotoxigenic Escherichia coli F4 type piglet diarrhea resistance and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Source of experimental animals

[0030] Collected from the Changping Experimental Pig Farm of the Chinese Academy of Agricultural Sciences.

[0031] There are 79 Landrace pigs, 203 Large White pigs, and 86 Songliao black pigs, including 301 piglets, a total of 348.

[0032] 2. Adhesion phenotype determination

[0033] After 35 days of age, the small intestinal epithelial tissue was slaughtered and collected for the adhesion test of Escherichia coli F4, which were divided into two types: adherent and non-adherent.

[0034] 3. Genomic DNA extraction

[0035] Pig ear tissue samples were collected, placed in centrifuge tubes filled with 70% alcohol, and stored in a -20°C refrigerator for later use.

[0036] Use the animal blood / tissue genomic DNA extraction kit to extract genomic DNA, the required reagents include:

[0037] Lysate laboratory equipment

[0038] Proteinase K (Germany MERCK Biotechnology Co., Ltd.)

[0039] Blood / cell / tissue genomic DNA extraction kit...

Embodiment 2

[0062] This example is the verification of the SNP site g.145109683 C / T obtained in Example 1 in Landrace, Large White and Songliao black pig populations.

[0063] 1. Extraction of pig genomic DNA

[0064] The ear tissue samples of three breeds of pigs with adhesion records were collected, placed in centrifuge tubes filled with 70% alcohol, and stored in a -20°C refrigerator for later use. Genomic DNA of pig ear tissue was extracted by the above method, and after quality and concentration detection, the concentration was diluted to 50 ng / μL and stored at -20°C for future use.

[0065] 2. PCR amplification and sequencing of the target fragment

[0066] Use the extracted pig DNA as a template, and perform PCR amplification according to the designed primers: the PCR amplification reaction system is: 1.0 μL of DNA template, 1.0 μL of primers shown in SEQ ID NO: 2 and SEQ ID NO: 3, Taq enzyme reagent 0.125 μL, double distilled water 17.38 μL; The reaction program of PCR amplifica...

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Abstract

The invention belongs to the technical field of molecular biology and discloses a SNP marker related to diarrhea resistance of enterotoxin-producing Escherichia coli F4 type piglets, a detection method and application thereof. The SNP marker is located on the nucleotide sequence of the integrin receptor protein ITGB5 gene on pig chromosome 13, and the site of the SNP marker is the g.145109683 nucleotide on pig chromosome 13 of the International Porcine Genome Version 10.2 Reference Sequence acid site, and has a C/T polymorphism, the SNP marker is extremely significantly correlated with diarrhea resistance of enterotoxigenic Escherichia coli F4 piglets. The SNP marker provided by the present invention is related to the diarrhea resistance of enterotoxigenic Escherichia coli F4 type piglets, and the diarrhea-resistant pig strain of enterotoxigenic Escherichia coli F4 type can be screened by identifying the SNP marker, and the obtained diarrhea-resistant strain has important Economic benefits and social value.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a SNP marker related to diarrhea resistance of enterotoxigenic Escherichia coli F4 type piglets, a detection method and application thereof. Background technique [0002] In current animal husbandry production, infectious diseases of livestock and poultry are not only a constraint factor for the development of animal husbandry, but also a serious threat to human food safety and even human health. Drugs and vaccines, as the main means to prevent and control infectious diseases of livestock and poultry, have played a key role, but at the same time of drug prevention and control, the drug resistance and mutation of pathogenic microorganisms are also enhanced at the same time, making disease prevention and drug treatment more and more difficult. How to get rid of the excessive dependence on drugs in animal husbandry production, reduce the use of antibiotics, and essentially i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 刘杨
Owner NANJING AGRICULTURAL UNIVERSITY
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