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SNP molecular marker associated with pig fat deposition, and applications thereof

A molecular marker, pig fat technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the lack of major genes and molecular markers, substantial progress, actual identification of major genes, Regulatory pathways and their applications are complex

Active Publication Date: 2015-11-18
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

People used the candidate gene method to screen some enzyme genes, hormone and receptor genes, cytokines and binding protein genes in the fat metabolism process related to pig carcass traits, but with the genotypes and QTLs detected by the effect significance The actual identification of main effect genes, regulatory pathways and their applications are becoming more and more complex, with little substantial progress
However, at present, in the practice of pig molecular breeding, there is still a lack of major genes and molecular markers with clear functions and significant effects

Method used

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  • SNP molecular marker associated with pig fat deposition, and applications thereof
  • SNP molecular marker associated with pig fat deposition, and applications thereof
  • SNP molecular marker associated with pig fat deposition, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The ear tissue samples of the Chinese local breed "Diannan small-eared pig" (collected from the Diannan small-eared pig resource protection farm in Xishuangbanna, Yunnan Province) and the introduced breed "Darkie pig" (collected from Anhui Kexin Pig Breeding Co., Ltd.) were selected. Genomic DNA was extracted by the phenol / chloroform method.

[0036] Design the following primers (as shown in the sequence listing SEQIDNO.2 and SEQIDNO.3):

[0037] Forward primer F: 5'-ACCTGACCTCCAGCTCTCAAAG-3'

[0038] Reverse primer R: 5'-GCAGTGATGGGGATGATGG-3'

[0039] Use the above primers to carry out PCR amplification on the genomic DNA of small-eared pigs and large grammes in southern Yunnan. PCR amplification uses a 20 μl reaction system: the amount of 2×PCRMix is ​​half of the volume of the PCR amplification reaction system, that is, 10 μl, and 100 ng of the pig genomic DNA to be detected , 10μmol / L forward and reverse primers 0.5μl each, and finally use ddH 2 Add O to 20 μl a...

Embodiment 2

[0043] Establishment of a sequencing method for genotype detection at A-931G site of porcine FNDC5 gene.

[0044] In order to detect the genotype of the A-931G locus of the pig FNDC5 gene quickly and conveniently, the genomic DNA of the pig is subjected to PCR amplification with the primer sequence in Example 1, and the PCR reaction system is a 20 μl system: 2×PCRMix dosage is the PCR amplification reaction Half of the system volume is 10 μl, 100ng of the porcine genomic DNA to be detected, 10μmol / L forward and reverse primers 0.5μl each, and finally use ddH 2Add O to 20 μl and mix well to obtain the PCR amplification reaction system. The PCR amplification reaction system was pre-denatured at 94-95°C for 5 minutes; 36 cycles were performed as follows: pre-denaturation at 94-95°C for 30 s, annealing at 60°C for 30 s, extension at 72°C for 40 s, and extension at 72°C for 7 min to obtain the PCR product.

[0045] The PCR amplification product was electrophoresed on agarose gel w...

Embodiment 3

[0047] The genotype distribution detection of the SNP site of the present invention in different pig groups.

[0048] The ear tissue samples of Tibetan pigs (Bujiangda County, Nyingchi, Tibet), Diannan small-eared pigs (Xishuangbanna, Yunnan), Yorkshire pigs (Anhui Kexin Pig Breeding Co., Ltd.) and Landrace pigs (Beijing Zhongshun Jingsheng Breeding Co., Ltd.) were collected and tested with phenol / chloroform method to extract pig individual genomic DNA samples.

[0049] The genotypes of the A-931G loci of the FNDC5 gene of the individuals of the four varieties were determined by amplified PCR product sequencing technology, and the detection results are shown in Table 1.

[0050] Table 1 Genotype distribution of pig FNDC5 gene A-931G locus

[0051]

[0052] Both Tibetan pigs and Diannan small-eared pigs belong to Chinese local pig breeds with slow growth rate, and their fat deposition ability is stronger than that of imported breed pigs, which belong to fat pigs. It can b...

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Abstract

The present invention relates to a SNP molecular marker associated with pig fat deposition, wherein the SNP molecular marker is located at 931bp on the upstream of the initiation codon of the pig FNDC5 gene 5' flanking region, and is recorded as A-931G. According to the present invention, the FNDC5 gene is adopted as the candidate gene affecting the pig fat deposition trait, the different SNP sites among different types of pig populations are obtained by using the PCR sequencing technology, the rapid detection method is established, the relevance between the SNP site and the pig fat deposition capability among the varieties and in the species is identified, and the SNP site affecting the pig fat deposition trait and the genotype are obtained.

Description

technical field [0001] The invention relates to a genotype detection method, in particular to a SNP molecular marker related to pig fat deposition and application thereof. Background technique [0002] Fat is one of the main components of animal body, a necessary substance to maintain normal life activities, and an important factor affecting the quality of pig carcass and meat flavor. Most local pig breeds in China are high-fat, called fat pigs; imported pig breeds have a high lean meat rate, called lean meat pigs. Genetic differences in carcass fat composition of pig breeds involve many genes and complex gene regulation, which has always been one of the difficult problems in the field of genetics and breeding. Metabolic diseases such as type 2 diabetes, obesity, fatty liver disease, hypertension, dyslipidemia, and atherosclerosis caused by excessive deposition of adipose tissue in the human body are threatening human health. Therefore, the regulation of fat metabolism has...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 张浩王志秀李庆岗商鹏张博
Owner CHINA AGRI UNIV
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