Method for identifying meat production traits of sheep and special primer thereof

A technology for sheep and traits, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., to achieve high application value, increase meat production, and high accuracy

Active Publication Date: 2019-01-18
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although our country is a big sheep-raising country in the world, our country is not a strong sheep-raising country. Therefore, the production capacity of mutton in our country needs to be improved

Method used

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  • Method for identifying meat production traits of sheep and special primer thereof
  • Method for identifying meat production traits of sheep and special primer thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Determination of embodiment 1 SNP site

[0021] 1. Selection of experimental materials

[0022] Collect 40 muscle tissues of Hu sheep and 39 muscle tissues of Suffolk sheep (Xinao Animal Husbandry Co., Ltd., Manas County, Changji, Xinjiang), and extract genomic DNA respectively, and adjust the DNA concentration of each sample to 100ng / μL, a total of 40 DNA samples of Hu sheep and 39 DNA samples of Suffolk sheep were obtained, which were stored at -80°C for future use.

[0023] 2. Design and synthesis of primers

[0024] Since the mature sequence of the sheep miR-1 gene has not been published, the mature sequence of the bovine miR-1 gene (MIMAT0009214, ID: bta-miR-1) was retrieved from the miRBase database (http: / / www.mirbase.org / ) , and compared with the NCBI sheep genome database (http: / / www.ncbi.nlm.nih.gov), and intercepted the 200bp sequences on the left and right flanks, and designed a pair of primers (primer combination) to clone sheep miR-1 Precursor gene, the...

Embodiment example 2m

[0035] Implementation case 2 Establishment of asymmetric PCR-SSCP detection method for miR-1 precursor gene

[0036] 1. PCR amplification

[0037] The genomic DNA of the sheep to be tested obtained in step 1 was used as a template, and U and D were used as primers to carry out PCR amplification to obtain PCR amplification products. PCR amplification system: 1 μL of 100ng genomic DNA, 10 μL of 2×Taq PCR MasterMix, 0.5 μL of forward primer (concentration of 10 μmol / L), 0.5 μL of reverse primer (concentration of 10 μmol / L), add ddH2O to 20 μL. PCR amplification program: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s; annealing at 60°C for 30 s; extension at 72°C for 50 s, a total of 30 cycles; final extension at 72°C for 10 min, detected by 1% agarose gel electrophoresis.

[0038] 2. Asymmetric PCR amplification conditions

[0039] The second PCR method is used, that is, the first PCR amplification is performed with primers of equal concentration to amplify t...

Embodiment example 3

[0045] Implementation Case 3 Correlation Analysis of Sheep miR-1 Precursor Gene g.53971777A>G Polymorphism and Meat Production-related Traits of Sheep

[0046] In order to determine whether the g.53971777A>G polymorphism site is related to the meat production of sheep, 40 Suffolk sheep and 39 Hu sheep were used as experimental materials, their genotypes were determined, and the relationship between the gene mutation site and the meat production Quantitative correlation analysis. The genotype and allele frequency of the g.53971777A>G polymorphism of the sheep miR-1 precursor gene in the population were calculated by SPSS13.0 software, and the chi-square test was used to analyze whether the genotype in each population was in the Hardy- Weinberg balance. The results are shown in Table 1.

[0047] The distribution and Hardy-Weinberg equilibrium of the gene frequency and genotype frequency of the SNP site of miR-1 gene in various sheep breeds

[0048]

[0049] Note: The numbe...

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Abstract

The invention discloses a method for assisting identification of meat production related traits of sheep and application thereof. The invention provides a method for assisting identification of meat-producing related traits of sheep, which comprises the following steps of: adopting specific primers to carry out PCR amplification on genomic DNA of sheep to be tested, utilizing DNA sequencing and asymmetric PCR; and using the specific primers to amplify the genomic DNA of sheep to be tested. DNA polymorphism detection and rapid typing by SSCP, The 209th nucleotide from 5 'end of PCR amplified product was A or G, and the genotype of sheep was GG or AA. There was a certain correlation between the SNP locus and the meat-producing traits of sheep. The meat-producing performance of sheep with GGgenotype was higher than that of sheep with AA genotype. The invention can be used for early screening of sheep meat production, effectively increasing the meat production of sheep breeding, reducingbreeding cost, and improving the economic benefit of sheep breeding. The method of the invention is simple in operation, low in cost and high in accuracy, and will have important significance for sheep breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method and application for assisting in identifying traits related to sheep meat production. Background technique [0002] Sheep play an important role in both agricultural production and people's life, and are important livestock animals. Over the years, the number of sheep and goats in my country, the production of mutton, sheepskin, sheep casings and cashmere have ranked first in the world. At the same time, the price of domestic mutton has always been higher than that of pork, beef and chicken, and has been rising all the way. With the improvement of people's living standards, the types of meat on the table are also increasing, and people are no longer simply satisfied with pork and other meats with high fat content and high cholesterol content. The protein and essential amino acid content of sheep meat is higher than that of pork, chicken and beef, the fat conten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/124
Inventor 胡圣伟倪伟张翔宇李村院李晓悦
Owner SHIHEZI UNIVERSITY
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