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SNP molecular marker related to pig carcass trait and application thereof

A technology of molecular markers and carcasses, which is applied in the direction of recombinant DNA technology, measurement/testing of microorganisms, DNA/RNA fragments, etc., and can solve the problem of few molecular markers

Inactive Publication Date: 2015-02-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are few studies on molecular markers for pig carcass traits. The SNP molecular markers for body size traits found in the coding region of the pig COL2A1 gene in the present invention are found for the first time.

Method used

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  • SNP molecular marker related to pig carcass trait and application thereof
  • SNP molecular marker related to pig carcass trait and application thereof
  • SNP molecular marker related to pig carcass trait and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Pig genomic DNA extraction (the sample is a purebred large white pig, a conventional breed provided by the experimental pig farm of Huazhong Agricultural University, Wuhan City, Hubei Province, China)

[0026] The DNA samples of the pig population (pure Large White pig) used in the present invention were all extracted using the genomic DNA kit (TIANamp Genomic DNA Kit) produced by Beijing Tiangen Biochemical Technology Co., Ltd. (operated according to the kit instructions), and the specific steps are as follows: Said:

[0027] (1) Put ear samples (tissues) from purebred large white pigs into 2ml EP tubes, wipe clean ophthalmic surgical scissors with alcohol cotton and cut them into a paste, then add 200μl buffer GA (included with the kit).

[0028] (2) Add 20 μl proteinase K solution (included with the kit), mix well and place in a 56°C water bath for overnight digestion.

[0029] (3) Add 200 μl buffer GB (included with the kit), mix thoroughly by inversion, and pla...

Embodiment 2

[0049] 1. Construct family and genotype scan

[0050] The test group of pigs (purebred Large White pigs) used for trait association analysis is the FCR project test group jointly established by the Animal Genetics, Breeding and Reproductive Molecular Biology Laboratory of Huazhong Agricultural University where the applicant is located and Pfizer Pharmaceutical Co., Ltd. A total of 236 pigs were fed in the experimental farm of Huazhong Agricultural University. They were slaughtered when their body weight reached 80-100kg. Samples were collected in 21 batches. Diagonal Length (CDL), Rib Number (RN), etc. Genomic DNA of purebred Large White pigs was extracted from individual ear margin tissue samples of each purebred Large White pig by the Laboratory of Animal Genetics, Breeding and Reproductive Molecular Biology, where Huazhong Agricultural University is located (see Example 1 for the preparation method), and placed in- Store at 20°C for later use.

[0051] The DNA samples of ...

Embodiment 3

[0060] Example 3 Differential detection of pig COL2A1 gene expression in individuals with different carcass lengths

[0061] 1. Select individuals to extract RNA.

[0062] Individuals whose genotypes at the rs80827825 locus were GG and AA were selected from the population in the above-mentioned Examples 1-2, and total RNA of cartilage tissue was extracted (the preparation method is as described below). The carcass straight length and oblique length of each individual were measured, and the results are shown in Table 2.

[0063] Table 2 Col2A1 gene expression QPCR analysis individual information

[0064]

[0065] Total RNA was extracted using TRIZOL from Invitrogen Reagent kit extraction, the specific operation steps are as follows:

[0066] (1) Take out the preserved purebred large white pig cartilage tissue sample, put it into a mortar filled with liquid nitrogen, and grind it into powder, weigh 0.05-0.1g of the powder into a 1.5mL EP tube, add 1ml TRIZOL (solution), v...

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Abstract

The invention belongs to technical field of preparations of livestock molecular markers, and particularly to preparation and application of an SNP molecular marker related to a pig carcass trait. The preparation comprises the following steps: cloning in pig Col2A1 gene to obtain a gene segment used as the molecular marker and related to the pig carcass trait, wherein the nucleotide sequence is as shown in a sequence table of SEQ ID NO:1, and SmaI-RFLP polymorphism is caused by the base substitution of A276-G276 in a 276bp point of the SEQ ID NO:1. The invention also discloses a method for the preparation of the molecular marker related to the pig carcass trait and the application of the prepared molecular marker in the investigation of polymorphism of the pig carcass trait to provide a new mark for marker assisted selection of a pig.

Description

technical field [0001] The invention belongs to the technical field of livestock molecular marker preparation, and in particular relates to a method for predicting pig carcass traits by using COL2A1 gene single nucleotide polymorphism and molecular markers thereof. The method is related to SNP molecular markers related to pig carcass traits. Background technique [0002] Molecular marker-assisted selection is an important application of genetic engineering in modern livestock breeding. The method of molecular markers can not only shorten the breeding time limit, but also greatly reduce the human and material resource consumption of breeding; The application potential in animal breeding has been greatly improved (Lu Shaoxiong, Wu Changxin., 2000). Molecular markers for assisted selection include protein markers, microsatellite markers, SNP markers, etc. Single nucleotide polymorphism (single nucleotide polymorphism, SNP) mainly refers to the DNA sequence polymorphism caused ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 赵书红牛丽莉袁俊董谦
Owner HUAZHONG AGRI UNIV
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