SNP molecular marker related with pig growth velocity and application thereof

A molecular marker and growth rate technology, applied in the field of molecular genetics, can solve the problems of lack of functional molecular markers, and achieve the effects of shortening the generation interval, low cost, and excellent economic value

Active Publication Date: 2019-05-28
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of molecular markers with clear functions, significant effects, and direct application in breeding.

Method used

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  • SNP molecular marker related with pig growth velocity and application thereof
  • SNP molecular marker related with pig growth velocity and application thereof
  • SNP molecular marker related with pig growth velocity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Expression Analysis of TRPC1 Gene in Example 1 Pig Longissimus Dorsi

[0034] The Chinese local breeds "Tibetan Pig" (collected from the teaching practice ranch of Tibet Agriculture and Animal Husbandry College), "Wujin Pig" (collected from the teaching practice ranch of Tibet Agriculture and Animal Husbandry College) and the imported breed "Darkie Pig" (collected from Anhui Pig Breeding Co., Ltd.) Longissimus dorsi muscle tissue samples at the age of 6 months, using the traditional method of Trizol to extract tissue RNA, and reverse transcribed into cDNA.

[0035] Download the porcine TRPC1 gene mRNA sequence (accession number NM_001145751.1) from NCBI, and use the software PrimerPremier 5.0 to design the following primers:

[0036] Forward primer F: 5'-CATCCAAAGGCAAGGTTA-3'

[0037] Reverse primer R: 5'-AAGTCCGAAAGCCAAGTA-3'

[0038] Use the above primers to carry out real-time fluorescence quantitative PCR amplification on the cDNA of Tibetan pig, Wujin pig and Yor...

Embodiment 2

[0040] Example 2 Screening of pig TRPC1 gene SNP sites

[0041] The Chinese local breeds "Tibetan Pig" (collected from the teaching practice ranch of Tibet Agriculture and Animal Husbandry College), "Wujin Pig" (collected from the teaching practice ranch of Tibet Agriculture and Animal Husbandry College) and the imported breed "Darkie Pig" (collected from Anhui Genomic DNA was extracted from ear tissue samples from Pig Breeding Co., Ltd. using the phenol / chloroform method.

[0042] Download the porcine TRPC1 gene sequence (accession number NC_010455.5) from NCBI, and use the software Primer Premier5.0 to design four pairs of primers for the DNA sequence of the TRPC1 gene downloaded from NCBI. Primer 1 amplifies the 5' of porcine TRPC1 gene Regulatory region -2142bp~-1548bp, the length of the target amplified fragment is 595bp; primer 2 amplifies the 5' regulatory region of porcine TRPC1 gene -1702bp~-942bp, and the length of the target amplified fragment is 761bp; primer 3 amp...

Embodiment 3

[0055] Example 3 Detection of the regulatory activity of the SNP site segment promoter of porcine TRPC1 gene

[0056] Primers were designed according to the sequence of the porcine TRPC1 gene 5' regulatory region, and the primer sequences were:

[0057] Forward primer F: 5'-CTAGAGTTGTGATGGGTCTTC-3';

[0058] Reverse primer R: 5'-GCTCATTGTAAATCTGTGGC-3'

[0059] The primer amplifies the 5' regulatory region of porcine TRPC1 gene -2142bp~-1548bp, which contains two SNP sites, C-1763T and C-1604T. The length of the target amplified fragment is 595bp, and the nucleotide sequence is as shown in SEQ ID NO.1 shown.

[0060] Using pig genomic DNA of different genotypes as a template, the PCR reaction was used to amplify, and the C-1763T and C-1604T sites in the 5' regulatory region of the TRPC1 gene were obtained as C / C and T / T haplotype sequences, respectively. Denote as CC and TT. The amplified fragments were digested, purified, and ligated to form recombinant pGL3-basic-TRPC1, ...

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Abstract

The invention provides an SNP molecular marker related with pig growth velocity and application thereof. According to the SNP molecular marker disclosed by the invention, a TRPC1 gene is utilized as acandidate gene which can affect the pig growth velocity; a sequencing technique is utilized to screen and identify SNP loci related with the pig growth velocity to obtain 2 SNP loci which can affectthe pig growth velocity and a gene type effect; the 2 SNP loci are respectively arranged (595bp)th and (539bp)th positions of a sequence shown in SEQ ID NO.1; allelotype of the 2 SNP loci is C and T;furthermore, alleles of C and C and T and T of the 2 SNP loci are completely linked on a genome; thus, any SNP locus can be applied to breeding pig breed growth rate characters; furthermore, the SNP loci cannot be limited by ages, genders and breeds of pigs, can be applied to early breeding of the pigs and can quicken a breeding process.

Description

technical field [0001] The invention relates to the field of molecular genetics, in particular to a SNP molecular marker related to pig growth speed and the application of the molecular marker in breeding. Background technique [0002] Pork has been occupying more than 60% of my country's total meat consumption for a long time, and it is the main body of animal husbandry production. As people's awareness of environmental protection gradually increases, domestic environmental protection policies are gradually tightened, and the cost of pig farming is also gradually rising; faster growth speed corresponds to high production efficiency, so growth speed is an important target trait in pig genetic improvement. However, the growth rate of pig breeds is regulated by multiple transcription factors and epigenetics, which has always been one of the difficult problems in the field of genetics and breeding. Traditional breeding mainly uses phenotypes to infer genotypes and estimate bre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
Inventor 张浩付玉田小龙张戌园张博张雅文
Owner CHINA AGRI UNIV
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