Molecular marker related to weaning weight and anti-diarrheal diseases of piglets and application of molecular marker in pig breeding

A piglet weaning and molecular marker technology, applied in the field of molecular genetics, to achieve the effect of improving weaning weight and resistance to diarrheal disease, increasing weaning weight of piglets, and improving resistance to diarrheal disease

Active Publication Date: 2017-03-22
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The disease resistance traits and weaning weight of pigs are complex traits controlled by multiple genes. At present, the research on molecular markers of these two traits has achieved certain results, but the number of markers obtained still cannot meet the needs of pig breeding. Continuously develop and develop new molecular markers

Method used

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  • Molecular marker related to weaning weight and anti-diarrheal diseases of piglets and application of molecular marker in pig breeding
  • Molecular marker related to weaning weight and anti-diarrheal diseases of piglets and application of molecular marker in pig breeding
  • Molecular marker related to weaning weight and anti-diarrheal diseases of piglets and application of molecular marker in pig breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Obtaining of molecular markers related to piglet resistance to diarrheal disease and weaning weight:

[0050] 1) Primer design and partial DNA sequence amplification of porcine IL6 gene

[0051] Download porcine IL6 gene mRNA sequence (GeneBank accession number: NM_214399.1) and genome sequence (GeneBank accession number: NC_010451.3 REGION: complement (101081042..101085451) GPC_000000591, shown in SEQ ID NO.1) from NCBI. The two sequences were compared, and the downloaded sequence was (NC_010451.3REGION:complement(101081042..101085451)GPC_000000591, shown in SEQ ID NO.1) model sequence, and Primer5 software was used to design primer IL6-3 for amplifying IL6 Part of the gene Inron2, Intron3 and the complete Exon3 region, primer IL6-5 was designed to amplify part of the Intron4 and part of the Exon5 region of the IL6 gene, all primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., and the primer sequences are listed in Table 1 below Show:

...

Embodiment 2

[0059] Embodiment 2: the detection method of HpaII site, its steps are as follows:

[0060] 1) Primer sequence

[0061] The primers used for PCR amplification are IL6-3 primers, and the primer sequences are as follows

[0062] IL6-3F: 5'TGCGAATGGGTTCTGACT 3';

[0063] IL6-3R: 5'AATCGCTCTTCCCTCTGG 3'

[0064] 2) PCR reaction conditions

[0065] Use IL6-3 primers to amplify in porcine genomic DNA. The PCR reaction system is 25 μL, including 1 μL of template DNA, 0.5 nmol / μL of upstream and downstream primers, 12.5 μL of PCR mix (Bao Biology), and add deionized water to the total volume. 25 μL. The PCR program was as follows: pre-denaturation at 94°C for 4 min; then denaturation at 94°C for 35 s, annealing at 53°C for 35 s, and extension at 72°C for 40 s, a total of 35 cycles; finally, extension at 72°C for 8 min.

[0066] 3) HpaII PCR-RFLP detection

[0067] The total PCR product digestion reaction system is 10 μl, including 8.5 μl of PCR product, 0.5 μl (10 U / μl) of HpaII...

Embodiment 3

[0068] Embodiment 3: the detection method of DraI site, its steps are as follows:

[0069] 1) Primer sequence

[0070] The primers used for PCR amplification are IL6-5 primers, and the primer sequences are as follows

[0071] IL6-5F:5'CAAATGTGAACCTCCTCC 3'

[0072] IL6-5R:5'AACAACCTGGCTCTGAAA 3'

[0073] 2) PCR reaction

[0074] Use IL6-5 primers to amplify in porcine genomic DNA. The PCR reaction system is 25 μL, including 1 μL of template DNA, 0.5 nmol / μL of upstream and downstream primers, 12.5 μL of PCR mix (Bao Biology), and add deionized water to the total volume. 25 μL. The PCR program was as follows: pre-denaturation at 94°C for 4 min; then denaturation at 94°C for 35 s, annealing at 50°C for 35 s, and extension at 72°C for 40 s, a total of 35 cycles; finally, extension at 72°C for 8 min.

[0075] 3) DraI PCR-RFLP detection

[0076] The total PCR product digestion reaction system is 10 μl, including 8.5 μl of PCR product, 0.5 μl (10 U / μl) DraI endonuclease, and 1...

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Abstract

The invention discloses a molecular marker related to the weaning weight and anti-diarrheal diseases of piglets and application of the molecular marker in pig breeding. Two SNP loci related to the weaning weight and anti-diarrheal diseases of the piglets are found in the IL6 gene by inventors, wherein one of the SNP loci related to the weaning weight and anti-diarrheal diseases of the piglets is 1753T/C which is located at the 1753th nucleotide in the IL6 gene sequence, and the other SNP locus related to the weaning weight of the piglets is 3789A/T which is located at the 3789th nucleotide of the IL6 gene sequence. Based on the SNP loci, the invention also provides a PCR-RFLP molecular marker containing the SNP loci. The piglets can be genotyped by the PCR-RFLP molecular marker technology, so that the piglets with the high weaning weight and anti-diarrheal disease ability are kept, and the breeding process of pigs is accelerated.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and in particular relates to the preparation, detection method and application of molecular markers related to piglet weaning weight and resistance to diarrheal disease. Background technique [0002] Weaning weight of piglets is an important indicator of pig production performance. Studies have shown that the weaning weight of piglets is significantly related to the age at slaughter and daily gain. With the increase of weaning weight of piglets, the daily gain of piglets increases and the age at slaughter decreases. In the modern intensive production of live pigs, piglet diarrhea is one of the most common problems in piglet production. Diarrhea can lead to slow growth and development of piglets, and even dehydration death, which seriously restricts the economic benefits of pig farming. [0003] How to increase the weaning weight of piglets and reduce the incidence of diarrhea in piglets has alw...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/158
Inventor 牛步月邢桂玲査安东王希彪狄生伟
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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