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Pig multi-omics integrated precision breeding method

A technology of piglets and genotyping, applied in biochemical equipment and methods, recombinant DNA technology, DNA/RNA fragments, etc., can solve problems such as insufficient understanding and understanding, incomplete pig reproductive efficiency, etc., to speed up the breeding process and reduce costs , Ease of use

Active Publication Date: 2021-11-02
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current understanding and understanding of the pig genome is far from enough, and the genes that affect pig reproductive efficiency (including pig litter size and average fetal weight, etc.) are not yet fully grasped. Further research is necessary to meet the needs of pig breeding methods

Method used

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  • Pig multi-omics integrated precision breeding method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] The acquisition of embodiment 1 SNP molecular marker

[0084] 1. Experimental sample collection

[0085] 375 large white pigs from a pig farm in Shandong were used for SNP typing of HSD17B1 gene. Collect ear tissue samples one by one, put them in 75% alcohol and store them at -20°C for DNA extraction.

[0086] 2. HSD17B1 gene SNP typing

[0087] 2.1 Extraction and detection of pig genomic DNA

[0088] (1) Cut an appropriate amount of pig ear tissue, cut it into pieces and put it into a 1.5mL Axgen tube.

[0089] (2) Take a 50mL BD centrifuge tube, mix proteinase K with a final concentration of 0.4mg / mL and lysis buffer evenly, and add 0.5mL lysis solution to the 1.5mL centrifuge tube containing pig ear tissue for lysis.

[0090] (3) Place the centrifuge tubes in parallel and evenly on the shaking plate of the thermostatic hybridization furnace (seal the tube caps tightly to avoid liquid leakage). Place at 55°C for more than 6 hours (full mixing of the sample during...

Embodiment 2

[0125] Example 2 Detection of HSD17B1 gene expression

[0126] 1. Experimental sample collection

[0127]The large white pigs and Meishan pigs used to detect the expression of HSD17B1 gene came from the pig farm of Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Wuqing District, Tianjin. The ovaries of sows at 49 days of gestation were collected, all samples were stored in liquid nitrogen, and total RNA was extracted. Three individuals of each species were collected as biological replicates.

[0128] 2. Detection of HSD17B1 gene expression

[0129] 2.1 Extraction of total RNA from ovarian tissue

[0130] The glassware and tweezers used in the RNA extraction process must be dry-baked at 200°C for 4 hours to inactivate RNase. Eppendorf tubes and pipette tips must use RNase inactivation products. During the entire experiment, experimenters must wear masks and latex gloves.

[0131] (1) Sample collection: Place the colle...

Embodiment 3

[0157] Embodiment 3 detects the kit of relevant SNP site

[0158] The SNP site detection kit of the present invention comprises the following components:

[0159] (1), dNTP (2.5mM);

[0160] (2), 5×PCR buffer;

[0161] (3), FastPfuDNA polymerase (250U);

[0162] (4), SNaPshot mixed solution;

[0163] (5), MgSO 4 Solution (50mM);

[0164] (6), 6×DNA Loading Buffer;

[0165] (7), PCR amplification primer (P 1 and P 2 ): its nucleotide sequence is shown in SEQ ID No.1 and SEQ ID No.2;

[0166] (8), extension primer: its nucleotide sequence is shown in SEQ ID No.3;

[0167] (9), exonuclease (250U);

[0168] (10), shrimp-alkaline phosphatase (100U);

[0169] (11), purification buffer:;

[0170] (12), ddNTP;

[0171] (13), the positive control specimen of the homozygous mutation of the SNP site: a plasmid containing a G base artificially synthesized at the porcine rs323407415 site;

[0172] (14), the positive control specimen of the heterozygous mutation at the SNP sit...

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Abstract

Pig multi-omics integrated precision breeding method, the invention discloses a SNP locus related to pigleting traits, the SNP locus is located at position 323407415 of the pig HSD17B1 gene, and the two alleles of the SNP locus are A or G; the three genotypes corresponding to the SNP locus are AA, GG and AG respectively, pigs with genotype AG and GG have higher litter size and litter weight than pigs with genotype AA . On the basis of the above SNP loci, the genetic progress of pigs with high production efficiency can be accelerated by selecting the genotype.

Description

technical field [0001] The invention relates to a multi-omics integrated precise breeding method for pigs, in particular to a multi-omics integrated precise breeding method for improving the reproductive efficiency of pigs. Background technique [0002] my country's pig breeding industry has a long history, and the pig breed resources are abundant and widely distributed, with huge utilization potential and broad development prospects. The production of breeding pigs is the foundation of the pig industry. Only through scientific feeding and management can the breeding pigs breed a large number of high-quality piglets and improve the economic benefits of the pig industry. [0003] With the continuous and rapid development of my country's economy, people's living standards are gradually improving, and the demand for pork is also increasing. Therefore, people are also paying more and more attention to how to improve the reproductive performance of pigs. Reproductive performance...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 李奎周荣杨亚岚唐中林刘颖李文通敖红牟玉莲
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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