Method for detecting fat coverage rate of Simmental cattle carcass by using DLK1 gene markers

Abstract

The invention discloses a method for detecting a fat coverage rate of Simmental cattle carcass by using DLK1 (Dalta Like non-canonical Notch ligand 1) gene markers. The method comprises the followingsteps: designing a pair of primers by using a DNA sequence of DLK1 gene of a Chinese Simmental cattle; performing strict PCR amplification and electrophoresis of samples on a PCR reaction system; screening and finding out two SNP sites by sequencing; screening from a population to obtain a restriction endonuclease Msp I at a DLK1-478 C / T site, wherein an excision base defined by the restriction endonuclease is CCGG; and screening to obtain a restriction endonuclease Ase I at a DLK1-609 T / G site, wherein T allele and TT mutant genotype individuals carrying the DLK1-478 C / T site in the Chinese Simmental cattle population have relatively high fat coverage rates and relatively good marbling; and G allele and GG mutant genotype individuals carrying the DLK1-609 T / G site have relatively high fatcoverage rates. According to the invention, the two mutation sites of the DLK1 gene can be used as genetic markers for predicting the fat coverage rate and marbling traits of the Chinese Simmental cattle, and are used for early marker assisted selection of beef cattle.

Description

technical field

[0001] The invention discloses a method for detecting the fat coverage rate of a Simmental cattle carcass with a DLK1 gene marker, which uses the DLK1 gene marker to detect the fat coverage rate and marbling traits of Chinese Simmental cattle, and belongs to the technical field of detecting beef quality traits. Background technique

[0002] Chinese Simmental beef is bright red in color, fine in texture, full of elasticity, moderate in marble pattern, white or light yellow in fat color, high in hardness in fat texture, and 100% body surface fat coverage, which is difficult for ordinary beef to achieve. standard. From the perspective of genes, molecular genetic markers are used as the main means of research on beef quality traits, and quantitative traits controlled by micro-effect polygenes are screened to optimize the breeding of cattle herds.

[0003] Single nucleotide polymorphisms (Single Nucleotide Polymorphisms, abbreviated as SNP) refer to the variation...

Examples

Embodiment 1

[0022] The acquisition of DLK1 gene fragments in Chinese Simmental cattle and the establishment of polymorphism detection methods in functional regions:

[0023] 1.1 Test materials: 237 28-month-old Chinese Simmental bulls came from Baolongshan beef cattle fattening farm in Tongliao City, Inner Mongolia. Jugular vein blood collection, all blood samples were 10 mL / head, anticoagulated with ACD anticoagulant, and frozen at -20°C. Genomic DNA was extracted from blood samples with a blood genomic DNA extraction kit;

[0024] 1.2 Primer design and PCR amplification: Chinese Simmental cattle were selected as test materials. In order to detect as many SNPs sites as possible, fragments with higher SNP density were selected, and the following primer pairs were designed according to the selected bovine DLK1 gene sequence:

[0025] Forward primer F: 5'-TCCACAGTGGAGGCTACTAAG-3';

[0026] Reverse primer R: 5'-CTTGTCTCCTGACTTCCTAAG-3';

[0027] Carry out PCR amplification in the Chinese ...

Embodiment 2

[0036] The DLK1 gene SNPs genetic markers obtained by screening were used to calculate the genotype frequency and gene frequency.

[0037] The C allele frequency of DLK1-478 C / T locus in Chinese Simmental cattle population is 0.865, and CC is the dominant genotype; the T allele frequency of DLK1-609 T / G locus is 0.846, and TT is the dominant genotype .

[0038]Table 1. Genetic diversity of 2 SNPs of DLK1 gene in Chinese Simmental cattle population

[0039] SNP mapping

Embodiment 3

[0041] Association analysis of DLK1 molecular genetic markers with Chinese Simmental beef quality and carcass traits:

technical field

[0001] The invention discloses a method for detecting the fat coverage rate of a Simmental cattle carcass with a DLK1 gene marker, which uses the DLK1 gene marker to detect the fat coverage rate and marbling traits of Chinese Simmental cattle, and belongs to the technical field of detecting beef quality traits. Background technique

[0002] Chinese Simmental beef is bright red in color, fine in texture, full of elasticity, moderate in marble pattern, white or light yellow in fat color, high in hardness in fat texture, and 100% body surface fat coverage, which is difficult for ordinary beef to achieve. standard. From the perspective of genes, molecular genetic markers are used as the main means of research on beef quality traits, and quantitative traits controlled by micro-effect polygenes are screened to optimize the breeding of cattle herds.

[0003] Single nucleotide polymorphisms (Single Nucleotide Polymorphisms, abbreviated as SNP) refer to the variation...

Examples

Embodiment 1

[0022] The acquisition of DLK1 gene fragments in Chinese Simmental cattle and the establishment of polymorphism detection methods in functional regions:

[0023] 1.1 Test materials: 237 28-month-old Chinese Simmental bulls came from Baolongshan beef cattle fattening farm in Tongliao City, Inner Mongolia. Jugular vein blood collection, all blood samples were 10 mL / head, anticoagulated with ACD anticoagulant, and frozen at -20°C. Genomic DNA was extracted from blood samples with a blood genomic DNA extraction kit;

[0024] 1.2 Primer design and PCR amplification: Chinese Simmental cattle were selected as test materials. In order to detect as many SNPs sites as possible, fragments with higher SNP density were selected, and the following primer pairs were designed according to the selected bovine DLK1 gene sequence:

[0025] Forward primer F: 5'-TCCACAGTGGAGGCTACTAAG-3';

[0026] Reverse primer R: 5'-CTTGTCTCCTGACTTCCTAAG-3';

[0027] Carry out PCR amplification in the Chinese ...

Embodiment 2

[0036] The DLK1 gene SNPs genetic markers obtained by screening were used to calculate the genotype frequency and gene frequency.

[0037] The C allele frequency of DLK1-478 C / T locus in Chinese Simmental cattle population is 0.865, and CC is the dominant genotype; the T allele frequency of DLK1-609 T / G locus is 0.846, and TT is the dominant genotype .

[0038]Table 1. Genetic diversity of 2 SNPs of DLK1 gene in Chinese Simmental cattle population

[0039] SNP mapping

Embodiment 3

[0041] Association analysis of DLK1 molecular genetic markers with Chinese Simmental beef quality and carcass traits:

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