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Reagent for detecting single nucleotide polymorphism (SNP) locus genotype significantly related to Chinese Simmental beef quality traits

A quality trait, Simmental's technology, applied in recombinant DNA technology, microbial assay/inspection, DNA/RNA fragments, etc., can solve the problem of liver cancer cell migration inhibition and other issues

Active Publication Date: 2021-04-09
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After interfering with MAT2B gene expression, the migration of liver cancer cells was significantly inhibited

Method used

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  • Reagent for detecting single nucleotide polymorphism (SNP) locus genotype significantly related to Chinese Simmental beef quality traits
  • Reagent for detecting single nucleotide polymorphism (SNP) locus genotype significantly related to Chinese Simmental beef quality traits
  • Reagent for detecting single nucleotide polymorphism (SNP) locus genotype significantly related to Chinese Simmental beef quality traits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 PCR Amplification and Sequencing of Mixed Pond of Simmental Cattle in China

[0053] 1. Experimental method

[0054] (1) sample

[0055] Blood sample DNA was extracted from 127 Chinese Simmental cattle and stored in a -80°C refrigerator in our research group.

[0056] (2) DNA extraction

[0057] DNA was extracted from Chinese Simmental cattle blood samples using a blood genomic DNA extraction kit according to the instructions. The purity and concentration of DNA were determined using NanoDrop ND-2000 ultra-micro spectrophotometer (Thermo Fisher Scientific), and the quality of DNA was detected by agarose gel electrophoresis.

[0058] (3) Primer design

[0059] According to the bovine MAT2B gene sequence published in Ensembl (ENSBTAT00000016399.4), primers were designed using PrimerPremier 6.0 software, upstream sequence: 5'-GAGAAGGAAGTTTCGTGTTTGG3-' (SEQ ID NO.1); downstream sequence: 5'-ACAACCTGATGCTTTACTGGAA-3'( SEQ ID NO.2), and synthesized by Sangon Bi...

Embodiment 2

[0065] Embodiment 2 single sample PCR-RFLP detection and genotyping

[0066] 1. Experimental method

[0067] A single Chinese Simmental cattle blood genome DNA was used as a template for PCR amplification. Use 1.5% agarose gel electrophoresis to detect the PCR amplification product and confirm the length of the target fragment, and then use the restriction endonuclease TaqI-V2 to perform specific enzyme digestion detection on the remaining PCR product. The enzyme digestion system is 10 μL, including cutsmart buffer 1 μL , TaqI-V2 enzyme 0.2 μL, PCR product 5 μL, ddH 2 O 3.8 μL, digestion at 65°C for 30 min. The digested products were detected by 1.5% agarose gel electrophoresis to confirm the individual genotypes of different samples.

[0068] 2. Experimental results

[0069] A single Chinese Simmental cattle blood genome DNA was used as a template for PCR amplification using primers. Electrophoresis detection confirmed that the bands of each product were uniform, and the...

Embodiment 3

[0071] Example 3 Analysis of the relationship between quality traits and genotypes of Chinese Simmental beef

[0072] 1. Experimental method

[0073] Using SeqMan analysis, observe the overlapping peaks of mixed pool PCR sequencing; use Popgene32 software to calculate the gene frequency, genotype frequency, genetic heterozygosity (H), effective allele number (Ne) and polymorphic information content (PIC) of alleles ;Calculate the χ2 value; Use SPSS 23.0 software in the single-factor variance multiple comparison analysis to test the relationship between Chinese Simmental beef quality traits and genotypes. When p<0.05, it means that the difference is significant, p<0.01 when the difference is extremely significant. Results are expressed as mean ± standard error.

[0074] 2. Experimental results

[0075] The results are shown in Table 1. In the I2-2382 G>T locus, the dominant genotype is TT, and the dominant allele is T, with frequencies greater than 0.5. The heterozygosity (...

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Abstract

The invention discloses a reagent for detecting single nucleotide polymorphism (SNP) locus genotype significantly related to Chinese Simmental beef quality traits. The SNP locus is located in MAT2B gene intron 2, and has three genotypes of GG, GT and TT, and the genotypes of the SNP locus significantly affect beef quality. When the genotype of a sample is GG, the slaughter acidity and carcass depth of the individual are obviously higher than those of individuals of other genotypes; and when the genotype of the sample is GG, the kidney fat content and carcass chest depth of the individual are significantly higher than those of an individual with the genotype of TT, and the waist beef thickness of the individual is significantly less than that of the individual with the genotype of TT. The invention lays a foundation for further constructing a core cattle herd with excellent high-quality beef quality traits, and provides a molecular theoretical basis for early breeding of high-quality beef cattle in the future.

Description

technical field [0001] The invention relates to the technical field of livestock molecular breeding, in particular to a reagent for detecting the genotype of a SNP site significantly related to the quality traits of Chinese Simmental beef. Background technique [0002] Chinese Simmental cattle are a large breed of milk and meat service breeds independently bred in my country after being introduced from abroad in the 1950s. Because of their strong physique, strong adaptability, and excellent milk, meat and service performance, they have become the most domestically raised and widely distributed breeds. The most extensive variety is the leading variety in my country's beef cattle industry. [0003] Beef is an indispensable meat variety in our daily diet, and its high nutritional value is recognized by the public. Improving the production efficiency of the beef cattle industry and producing more safe and high-quality beef products has become an important issue restricting the de...

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 姜平阮梦茹赵志辉
Owner GUANGDONG OCEAN UNIVERSITY
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