Identification method of PPARD major gene, establishment of molecular breeding method and application thereof

A main effect gene and genotype technology, applied in the field of animal genetics and breeding, can solve problems such as phenotype separation

Inactive Publication Date: 2010-08-18
JIANGXI AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

Since the ancestor white Duroc is a lean male parent line widely used in commercial production, and the Erhualian pig is a local Chinese pig breed known f

Method used

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  • Identification method of PPARD major gene, establishment of molecular breeding method and application thereof
  • Identification method of PPARD major gene, establishment of molecular breeding method and application thereof
  • Identification method of PPARD major gene, establishment of molecular breeding method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Example 1: Effects of PPARD G32E on ear traits, fat deposition, head weight and carcass length in the white Duroc × Erhualian resource population.

[0098] (1) Genotype determination of PPARD G32E:

[0099] Design primer pair F2 and R2 (F2: 5'-CGG CTG TTT TAC AGG AAG GA-3'; R2: 5'-CTG CAC TCA GAC CCA GAT GA-3') to amplify the DNA fragment containing PPARD G32E, design and synthesize Another extension primer for SNAPshot reaction (5'-TTT TTT TTT TGC TGG AGGGAA GCG AGT GCT CTG GT-3'). In a 15μl PCR reaction system using primers F2 and R2 to amplify porcine genomic DNA, including 40ng porcine genomic DNA, 35mM MgCl 2 , 20mM dNTP, 2pmol each of the forward and reverse primers, 2 units of Taq enzyme and 1×PCR buffer (Shanghai Bioengineering Company, China). Amplification conditions were: 94°C for 3 min; 94°C for 30 s, 67°C for 30 s, 72°C for 40 s, a total of 30 cycles; finally extended at 72°C for 10 min. PCR products were detected by 2% agarose gel electrophoresis and ph...

Embodiment 2

[0107] Example 2: Effects of PPARD G32E on ear area in 6 distantly related populations.

[0108] The applicant collected ear tissue samples from 574 adult sows with more than 2 litters in 6 distant populations including Erhualian pig, Hang pig, Yushan black pig, Suzhong pig, Sujiang pig and Sutai pig, and extracted Genomic DNA. The ear area of ​​the tested individuals was measured by the following method: place a standard ruler parallel to the pig's ear, take a high-resolution photo of the pig's ear through vertical focusing, use the standard ruler as a reference, and use the LaicaQwin area measurement software to calculate Calculate the ear area of ​​each individual.

[0109] According to the method of step 1 in Example 1, using the genomic DNA of the above-mentioned distant population individuals as a template, PCR amplification was carried out under the guidance of the primer pair F2 and R2, and 5'-TTT TTT TTT TGC TGG was detected using the SNAPshot technology of ABI compa...

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Abstract

The invention relates to an identification method of PPARD major gene, establishment of a molecular breeding method and application in the genetic improvement of breeding pigs. In the invention, by adopting the white Duroc * Erhualian Intercross and the Chinese and exotic breeding pigs of distant population, the whole-genome scan and linkage mapping, the IBD fine mapping of the target region based on a high-density SNP mark and the selection effect analysis of the distant population are carried out, and then the major gene, i.e., PPARD for deciding the traits of pig ears, fat deposition, weight of pig head and carcass traits, and the mutation site of key cause, i.e., G32E are identified. The sites are detected by the modern molecular biotechnology; by adopting the gene-assisted selection method, the beneficial gene-typed individuals are selected for the breeding pigs, thereby the breeding improvement efficiency of the traits of pig ears, fat deposition, weight of pig head and carcass traits are remarkably improved. The invention further discloses an identification course of the gene and the cause mutation site and a method for breeding the traits of pig ears, fat deposition, weight of pig head and carcass traits by using the gene marker of the invention.

Description

technical field [0001] The invention relates to the field of animal genetics and breeding, in particular to the identification of a main gene affecting pig ear traits, fat deposition, body length and head weight, the establishment of a molecular breeding method and the application of the gene in genetic improvement of breeding pigs. Background technique [0002] China is a big pig raising country and the country with the richest pig breeding resources. It has long domesticated wild boars into domestic pigs. Through long-term selection, many distinctive local breeds have been formed, and there are only 48 recorded in "Chinese Pig Breeds Chronicle" (Edited by Zhang Zhongge, Shanghai Science and Technology Press, 1986). Compared with foreign commercial pigs, Chinese local breeds have formed significantly different body appearance and germplasm characteristics in the long-term breeding history. Taking ear characteristics and body length as an example, pigs with large and droopi...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47C12Q1/68
Inventor 黄路生任军段艳宇乔瑞敏姚飞
Owner JIANGXI AGRICULTURAL UNIVERSITY
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