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SNP molecular markers for traceability on pig chromosome 6 and its application

A technology of molecular markers and chromosomes, applied in the direction of recombinant DNA technology, DNA / RNA fragments, etc., can solve the problems of indistinguishable individuals, molecular markers cannot be independent of each other, molecular marker linkage phenomenon, etc., to expand the detection range, improve detection coverage, The effect of improving accuracy

Active Publication Date: 2018-01-26
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using the existing 13 known SNP molecular markers as a combination of traceability markers, some individuals may not be able to distinguish when using this set of markers for traceability in highly selected pig populations
[0007] In addition, in the long-term breeding work, a large number of accumulated SNP markers are often concentrated on some chromosomes, while other chromosomes, such as chromosomes 5, 6, 8, 14, 15 and 18 The lack of SNP markers can easily lead to linkage between molecular markers, and the molecular markers cannot be independent of each other, reducing the detection range of traceability experiments, resulting in the actual detectable range being smaller than the expected detectable range

Method used

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  • SNP molecular markers for traceability on pig chromosome 6 and its application
  • SNP molecular markers for traceability on pig chromosome 6 and its application
  • SNP molecular markers for traceability on pig chromosome 6 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Search for Molecular Markers

[0029] (1) Primer design

[0030] Using the DNA sequence of pig chromosome 6 (Genbank: FN675082) as a template, design primers to isolate the DNA fragment of pigs (using the DNA of a Duroc pig as a template for PCR amplification), the primers are as follows:

[0031] Forward primer: 5'-AAATGAAGAACAGGCAAGG-3' (as shown in SEQ ID NO.2)

[0032] Reverse primer: 5'-CTTCCCAGTAAACTTAGGAGATA-3' (as shown in SEQ ID NO.3)

[0033] The total volume of the PCR reaction is 20μl, including about 100ng of porcine genomic DNA, containing 1×buffer (Promega Company), 1.5mmol / L MgCl 2 , the final concentration of dNTP (Shanghai Sangon Biotechnology Co., Ltd.) was 150 μmol / L, the final concentration of primers was 0.2 μmol / L, and 2U Taq DNA polymerase (Promega Company).

[0034] PCR amplification: 94°C for 4min, (94°C for 30s, 60°C for 30s, 72°C for 30s) cycled 30 times, and finally 72°C for 10min.

[0035] PCR reaction products were detected b...

Embodiment 2

[0046] Example 2 Distribution of alleles

[0047] (1) Design of test groups

[0048] Experimental group: collected Pietrain (21 heads), Shannon (17 heads), Dabai (43 heads), Dashen (52 heads), Changshen (16 heads), Dushen (23 heads), Pi Dashen (11 heads) DNA was extracted from the ear tissues of individuals of the head), Dachang Shen (25 heads), Du Dashen (17 heads) and Dupi Dashen (20 heads), totaling 245 DNA samples.

[0049] The purpose of the test population is to detect the distribution of SNP markers in different breeds.

[0050] (2) Genetic testing

[0051] Forward primer: 5'-AAATGAAGAACAGGCAAGG-3' (as shown in SEQ ID NO.2);

[0052] Reverse primer: 5'-CTTCCCAGTAAACTTAGGAGATA-3' (as shown in SEQ ID NO.3).

[0053] Amplify (amplification conditions are the same as in Example 1), and detect all individual genotypes of the test population with the same PCR-TaiI-RFLP method.

[0054] (3) Statistical analysis

[0055] Record all individual genotypes of the test populat...

Embodiment 3

[0059] Example 3 SNP molecular marker usability traceability test verification

[0060] The SNP molecular marker of the present invention has been preliminarily confirmed that it can be used in traceability markers. In order to ensure the usability in traceability markers, the present invention re-sampled in different locations, and existing 13 SNP molecular markers (as shown in Table 2 below) Shown) combined use (total 14 SNP molecular markers) to carry out usability traceability test verification, at the same time, use only the existing 13 SNP molecular markers to carry out traceability test verification as a comparison example.

[0061] Randomly collect 100 ear tissue samples and 100 muscle samples from individual pigs at Fuxing Slaughterhouse (every pig individual sample is collected at the same time as ear tissue samples and muscle samples), ear tissue samples are numbered E1-E100, and muscle samples are numbered M1-M100, respectively DNA from muscle samples and ear tissu...

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Abstract

The invention discloses a SNP molecular marker used for traceability on pig No. 6 chromosome and its application. The SNP molecular marker is obtained through NCBI database sequence alignment, and the SNP molecular marker is specifically recognized by a TaiI restriction endonuclease. A total of 550bp, there is a base substitution at the 362nd base, which is 362A or 362G, and its sequence is shown in SEQ ID NO.1. By analyzing the distribution of the alleles of the SNP molecular markers in the test populations including 10 pig breeds or strains, it was found that the allele frequencies of the SNP molecular markers of the present invention in different breeds or strains were close, polymorphic There are few differences in the distribution of allele frequencies between breeds or lines, and the heterozygosity is greater than 0.3. This SNP marker can be used for DNA traceability of pork products.

Description

technical field [0001] The invention belongs to the field of food safety, and relates to a SNP molecular marker used for traceability on pig No. 6 chromosome and a detection method thereof. Background technique [0002] With the development of society, the links in the food supply chain are constantly increasing. From farms, pastures to processing, packaging, storage, transportation and sales of food enterprises, there are food safety hazards in all links of food supply, and food safety problems often occur. occur. Therefore, it is necessary to carry out traceability management of meat products, and establish a traceability system for all links from the place of origin to the dining table, so as to provide consumers with accurate and detailed information about products, which is conducive to timely discovery of unsafe conditions in various links by producers and operators Hidden danger. Since the food safety crisis in the mid-1990s, the traceability of meat products has be...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6888
Inventor 吴潇唐雪明吕贝贝蒋玮王金斌武国干李鹏白蓝刘华赵凯
Owner SHANGHAI ACAD OF AGRI SCI
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