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
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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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