Multi-omics integration accurate pig breeding method

A technology for piglets and aspects, applied in the field of multi-omics integrated precision breeding of reproductive efficiency, can solve problems such as insufficient understanding and understanding, incomplete pig reproductive efficiency, etc., and achieve the effect of accelerating the breeding process, accelerating genetic progress, and promoting breeding selection

Inactive Publication Date: 2019-03-19
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 (inclu

Method used

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  • Multi-omics integration accurate pig breeding method
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  • Multi-omics integration accurate pig breeding method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1 Detection of HSD17B1 gene expression

[0088] 1. Collection of experimental samples

[0089] The large white pigs and Meishan pigs used to detect the expression of the HSD17B1 gene came from the pig farm of the 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, and the samples were stored in liquid nitrogen to extract total RNA. Three individuals of each species were collected as biological replicates.

[0090] 2. HSD17B1 gene expression detection

[0091] 2.1 Extraction of total RNA from ovarian tissue

[0092] The glassware and tweezers used in the RNA extraction process need to be dry-baked at 200°C for 4 hours to inactivate RNase. Eppendorf tubes and pipette tips need to use RNase inactivation products. The experimenters need to wear masks and latex gloves during the entire experiment.

[0093] (1) Sample collection: Place the ...

Embodiment 2

[0119] Example 2 Obtaining SNP molecular markers

[0120] 1. Collection of experimental samples

[0121] 375 large white pigs from a certain 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 at -20°C, and prepare DNA.

[0122] 2. SNP typing of HSD17B1 gene

[0123] 2.1 Extraction and detection of pig genomic DNA

[0124] (1) Cut an appropriate amount of pig ear tissue, cut it into pieces, and put it into a 1.5 mL Axgen tube.

[0125] (2) Take a 50 mL BD centrifuge tube, mix the proteinase K with a final concentration of 0.4 mg / mL and the lysis buffer uniformly, and add 0.5 mL of the lysis buffer to a 1.5 mL centrifuge tube containing pig ear tissue for lysis.

[0126] (3) The centrifuge tube is evenly placed on the shaking plate of the thermostatic hybridization furnace in parallel (seal the tube lid tightly to avoid liquid leakage). Store at 55°C for more than 6 hours (full mixing of samples during di...

Embodiment 3

[0161] Example 3 Kit for detecting relevant SNP sites

[0162] The SNP site detection kit of the present invention includes the following components:

[0163] (1), dNTP (2.5mM);

[0164] (2), 5×PCR buffer;

[0165] (3) FastPfuDNA polymerase (250U);

[0166] (4), SNaPshot mixture;

[0167] (5), MgSO 4 Solution (50mM);

[0168] (6), 6×DNA Loading Buffer;

[0169] (7), PCR amplification primers (P 1 And P 2 ): Its nucleotide sequence is shown in SEQ ID No. 4 and SEQ ID No. 5;

[0170] (8) Extension primer: its nucleotide sequence is shown in SEQ ID No. 6;

[0171] (9) Exonuclease (250U);

[0172] (10), shrimp-alkaline phosphatase (100U);

[0173] (11) Buffer for purification:;

[0174] (12), ddNTP;

[0175] (13) Positive control specimen for homozygous mutation of SNP site: a plasmid containing pig rs323407415 site artificially synthesized as G base;

[0176] (14) Positive control specimen of heterozygous mutation at SNP site: a mixture of two plasmids containing artificially synthesized A and G bas...

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Abstract

The invention discloses a method for identifying or auxiliary identifying of pig farrowing characters and relates to a multi-omics integration accurate pig breeding method. The pig farrowing characters are determined through determination of mRNA expression quality of a pig HSD17B1 gene. Particularly, by acquisition of total RNA of pig ovary tissue cells, reverse transcription and quantitative PCR, the mRNA expression quality of the pig HSD17B1 gene is determined, and pigs in low mRNA expression quality are high in litter size and/or litter weight. In addition, by adoption of the identification method and artificial selection, high-litter-size pig breeding can be implemented. The identification method and application thereof to breeding are independent of genome editing means, acceptance of people is improved, and new breakthrough in breeding pig breeding is realized.

Description

Technical field [0001] The invention relates to a multi-omics integrated precision breeding method for pigs, in particular to a multi-omics integrated precision breeding method for improving the reproductive efficiency of pigs. Background technique [0002] The pig industry in my country has a long history, with abundant pig breed resources and wide distribution, with huge utilization potential and broad development prospects. Breeding pig production is the foundation of the pig industry. Only through scientific breeding and management, can breeding pigs produce large numbers and 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 paying more and more attention to how to improve the reproductive performance of pigs. Reproductive performance is an important economic tr...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/178
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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