A SNP molecular marker affecting the number of thoracic vertebrae in sheep and its application

A molecular marker and molecular marker-assisted technology, applied in the field of molecular biotechnology and molecular markers, to achieve the effects of improving reproductive performance, rapid and accurate selection and improvement, and increasing core competitiveness

Active Publication Date: 2022-03-18
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Molecular markers affecting the number of thoracic vertebrae have not yet been identified at home and abroad

Method used

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  • A SNP molecular marker affecting the number of thoracic vertebrae in sheep and its application
  • A SNP molecular marker affecting the number of thoracic vertebrae in sheep and its application
  • A SNP molecular marker affecting the number of thoracic vertebrae in sheep and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Experimental animals

[0037] The experimental sheep group used in the present invention is 496 Ujimqin sheep selected from the grassland area of ​​East Ujimqin Banner in Inner Mongolia Autonomous Region.

[0038] 2. Sample collection

[0039] The muscle tissue of the slaughtered sheep was collected and soaked in ethanol with a volume fraction of 75%, and stored in a -20°C refrigerator for later use.

[0040] 3. Whole sheep genome 10× resequencing

[0041] Using the muscle tissue collected from each of the 496 sheep in step 1, the whole genome DNA was extracted by the standard phenol-chloroform method, and the DNA concentration and OD ratio of each sample were accurately determined by the Nanodrop2000 / 2000C nucleic acid and protein detector (OD260 / 280, OD260 / 230). Qualified DNA samples detected by the NanoDrop2000 / 2000C Nucleic Acid Protein Detector. Pipette 3 μL of the extracted DNA sample to be tested and mix it with 1.5 μL Loading Buffer, load the sample onto ...

Embodiment 2

[0051] Example 2 Target DNA sequence amplification and sequencing

[0052] (1) Primer design

[0053] The DNA sequence of SEQ ID NO: 1 on chromosome 7 of sheep was downloaded from the UCSC website (http: / / genome.ucsc.edu / ). And use primer design software primer premier 3.0 to design primers.

[0054] The DNA sequences of the designed primers are as follows:

[0055] P001-F: 5'-TGCAATTGGCTGTTTGAGTC-3';

[0056] P002-R: 5'-ACTTGCATCCCTTCAGCAGT-3';

[0057] (2) PCR amplification

[0058] Add 1 μL of DNA template, 10.5 μL of double distilled water, 12.5 μL of 2×Dream Taq Green PCRMaster mix (Thermofisher), and 0.5 μL of primers P001-F and P002-R into the 25 μL reaction system.

[0059] The PCR reaction conditions were: pre-denaturation at 95°C for 5 min; 30 cycles of denaturation at 95°C for 30 s, annealing at 60°C for 30 s, and extension at 72°C for 30 s; and extension at 72°C for 10 min.

[0060] (3) DNA sequence determination

[0061] DNA sequence sequencing identificati...

Embodiment 3

[0064] Example 3 SNP site 82509619G>A effect analysis of molecular markers

[0065] Through molecular marker-assisted selection and elimination of sheep with genotypes of GA and GG in the group, we have significantly increased the total number of thoracic vertebrae in the group, promoted the breeding process of sheep, increased the meat production performance of the sheep group, and driven the economy of the mutton sheep industry Benefits are maximized.

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Abstract

The invention belongs to the technical fields of molecular biotechnology and molecular markers, and specifically relates to a SNP molecular marker that affects the number of thoracic vertebrae in sheep and its application. The SNP site of the SNP molecular marker affecting the number of thoracic vertebrae in sheep corresponds to the G>A mutation at position 82509619 on chromosome 7 of the International Sheep Genome Version 4.0 Reference Sequence. By optimizing the dominant allele of the SNP, the invention can increase the frequency of the dominant allele generation by generation, improve the meat production performance of sheep, accelerate the progress of sheep genetic improvement, and effectively improve economic benefits.

Description

technical field [0001] The invention belongs to the technical fields of molecular biotechnology and molecular markers, and specifically relates to a SNP molecular marker that affects the number of thoracic vertebrae in sheep and its application. Background technique [0002] The mammalian spine is divided into five parts from front to back: cervical vertebra, thoracic vertebra, lumbar vertebra, sacral vertebra and caudal vertebra. The number of sternums in each part of most mammal species is fixed. There is a certain range of variation in the total number of thoracolumbar vertebrae in a small number of mammalian species, such as pigs, cattle, and sheep. The number of vertebrae is significantly correlated with carcass weight and carcass length, and has high heritability, for example, the heritability in pigs is 0.60-0.62. [0003] Sheep are the main livestock of grassland herdsmen, and are also known as the first of the "five grassland animals". Mutton and wool of sheep ar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/124
Inventor 张慧王小鹏黄思秀蔡更元黄敏杨丽娟刘冰冰刘少娟郑素梅余志强李德森李桂新蒋勇闯
Owner SOUTH CHINA AGRI UNIV
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