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Molecular biological method for breeding high-quality mutton sheep

A mutton sheep and molecular marker technology, which is applied in the field of molecular biology and molecular breeding to achieve the effect of speeding up the breeding process, promoting economic benefits and promoting improvement

Active Publication Date: 2020-11-24
黑龙江省农业科学院畜牧兽医分院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, no research has reported that there are molecular markers related to Dorper lamb meat quality on the PDK4 gene

Method used

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  • Molecular biological method for breeding high-quality mutton sheep
  • Molecular biological method for breeding high-quality mutton sheep
  • Molecular biological method for breeding high-quality mutton sheep

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Discovery of polymorphic sites in Dorper sheep PDK4 gene

[0023] The experimental subjects were selected from 200 healthy 10-month-old Dorper sheep in the applicant's own farm, and 2ml of blood was drawn from the jugular vein, anticoagulated, and stored at -20°C until use.

[0024] (1) Extraction of genomic DNA. Genomic DNA was extracted using the Blood Genomic DNA Extraction Kit from Tiangen Biochemical Technology Co., Ltd., and no tailing or smearing was detected by 1% agarose gel electrophoresis. The DNA was detected with an ultraviolet spectrophotometer, and the A260 / 280 ratio was in the range of 1.8-2.0, and the A260 / 230 ratio was in the range of 1.7-1.9, and the quality of the sample was qualified. Dilute the sample to 50ng / μl and freeze it at -20°C.

[0025] (2) Exon sequencing of PDK4 gene. 20 Dorper sheep DNA samples were randomly selected to construct pool DNA for exome sequencing. The genomic DNA sequence of the sheep (Ovis aries) PDK4 gene (ac...

Embodiment 2

[0027] Example 2 Genotype frequency and allele frequency of polymorphic loci

[0028] (1) The DNA fragment of exon 4 of PDK4 containing the mutation site was amplified by PCR. Primers were designed using Primer Premier 5.0 to amplify the DNA fragment containing the mutation site and the restriction endonuclease site analysis was performed. Upstream primer F: 5'-CATCAAAGTTCGAAACAGACACCA-3' (SEQ ID NO: 2); Downstream primer R: 5'-GCACAGCTTATATTTCTCCAGCCA-3' (SEQ ID NO: 3). PCR reaction system: 10 μl system contains 1 μl of template, 0.2 μl of upstream and downstream primers, 5 μl of Taq DNA polymerase, and milli-Q water for the rest; PCR reaction program: pre-denaturation at 94°C for 2 minutes, 35 cycles (denaturation at 94°C for 30s , annealed at 60°C for 30s, extended at 72°C for 30s), extended at 72°C for 5min, and stored at 4°C until use. The PCR product is a 233bp DNA fragment (SEQ ID NO: 4), wherein the 80th position is a single nucleotide mutation site (G>T). After ana...

Embodiment 3

[0035] Example 3 Correlation analysis between different genotypes and meat quality traits

[0036] (1) Determination of intramuscular fat content. Intramuscular fat is one of the main indicators of mutton meat quality. Within the normal range, intramuscular fat content is positively correlated with meat tenderness, taste flavor and juiciness. When a certain amount of fat is deposited between the muscle fibers, the marbling increases, and the tenderness and juiciness of the meat are improved. Determination of intramuscular fat content using the Soxhlet extraction method to detect the intramuscular fat content of the longest muscle of the sheep's back: (1) Grind 3-5g of the sample in liquid nitrogen to a fine powder, and put it into a weighed drying filter paper In the bag (W1), weigh the filter paper bag containing the wet sample (W2), after drying for 2 hours, weigh the filter paper bag containing the dry sample (W3); (2) use a Soxhlet extractor to extract with ether at room...

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Abstract

The invention belongs to the fields of molecular biology and molecular breeding, and concretely provides an SNP molecular marker related to mutton quality traits on a PDK4 gene of Dorper sheep, a detection method of the molecular marker and application of the method to improvement of mutton quality and breeding of high-quality mutton sheep varieties. The molecular marker is a polymorphic site at the 95th site of an exon 4 of a PDK4 gene and is marked as G95T. Statistical analysis shows that the molecular marker G95T is significantly related to mutton quality traits, especially the intramuscular fat content, the muscle fiber diameter and the water loss rate. Therefore, in the mutton sheep breeding process, fresh, tender and juicy GT type or TT type individuals can be selected in an auxiliary way through the SNP molecular marker, so that the high-quality mutton sheep breeding process is accelerated.

Description

technical field [0001] The invention belongs to the fields of molecular biology and molecular breeding. Specifically, the present invention provides a SNP molecular marker related to Dorper sheep PDK4 gene and meat quality, a detection method of the molecular marker and the application of the method in improving the meat quality level of mutton and breeding high-quality mutton sheep breeds. Background technique [0002] Meat quality-related traits have important economic value in mutton sheep breeding. The quality of meat is affected by internal and external factors such as genes, feeding, and breeding conditions. Meat production performance mainly includes indicators such as slaughter rate, ketone weight, fat content, etc. Meat quality also includes pH 24 , cooked meat rate, water loss rate, shear force, meat color, tenderness, muscle fiber diameter and other indicators. These indicators can directly reflect the quality of meat. The rapid development of DNA molecular ...

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 刘学峰李旭业海龙李信涛王洪宝张立春姜明生吴宪王嘉厚郭立宏郝彩虹吕雄杰孙继权杜军穆文利王锋尤海洋李莉郭文凯杨淑萍刘丽秋刘秀玲王佳王宇董扬张超
Owner 黑龙江省农业科学院畜牧兽医分院
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