Method for detecting chicken fatty character
A technology for fat and traits, which is applied in the field of detection of chicken fat traits using the single nucleotide polymorphism of the PGC-1α gene, can solve the problems of difficult abdominal fat traits and low efficiency of breeding methods, and achieve high accuracy and speed up Breeding process, simple operation effect
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Embodiment 1
[0018] Embodiment 1, establishment of PCR-SSCP detection method and determination of polymorphic sites
[0019] 1. PCR amplification
[0020] Select 2 laying hen breeds (Bailaihang layer and Nongda No. 3 grain-saving small laying hen), one broiler breed (Bai Luoke) and three Chinese local breeds (Dongxiang green-shell layer, silky silky and Beijing You Chicken) is the experimental material, and according to the cDNA sequence (GenBank accession number: AB170013) of chicken PGC-1α gene, primers are designed, and the sequence is as follows: F (forward primer): 5'-GACTGAGAATTCGTGGAGCA-3' (SEQ ID NO in the sequence listing: 1) R (reverse primer): 5'-ACCTTCCCTCCAAAACCAAC-3' (SEQ ID NO: 2 in the sequence listing)
[0021] Using the genomic DNA of the above five chicken species as templates, PCR amplification was carried out under the guidance of primers F and R. The 20 μl PCR reaction system was: chicken genomic DNA 100ng, 10×PCR amplification buffer 2.0 μL, dNTPs 200 μmol / L , 50ng...
Embodiment 2
[0026] Example 2, Detection of polymorphic distribution of molecular markers in different flocks
[0027] For 2 laying hen breeds (Bai Lai Hang layer and Nongda No. Chicken) The same method as in Example 1 was used to detect the G / A polymorphism of the 646th base of the PGC-1α gene cDNA from the 5' end, and the detection results are shown in Table 1. Among the tested chicken breeds, the gene frequency of the dominant G allele in the White Rock broiler chicken was 0.67, while the gene frequency of the dominant A allele in the White Leghorn laying hens was 0.82, while in Dongxiang green-shell eggs The frequencies of G allele and A allele in chicken, silky chicken and Beijing oil chicken were similar. x 2 The results of fitness test analysis showed that the distribution of genotypes in these populations was in a balanced state (P>0.05), indicating that the gene frequency was not affected by selection during the evolution process, and the results obtained were credible.
[0028...
Embodiment 3
[0029] Example 3. Correlation analysis between molecular markers and fat traits
[0030] In order to determine whether the G / A polymorphism of the 646th base (G646A) of the PGC-1α gene cDNA from the 5' end is related to the phenotypic differences of chickens, the F2 population (332 Only) is the test material, polymorphism detection is carried out with the same method as Example 1, and the correlation between different genotypes of chicken PGC-1α gene G646A and production traits is analyzed. SAS statistical analysis software GLM program was used to conduct single-marker analysis of variance. Genotype, reciprocal cross and batch were fixed effects. For carcass traits, body weight at slaughter was introduced into the model as a covariance for statistical analysis.
[0031] Results Among the 332 F2 individuals tested, there were 109 AA genotypes, 176 AB genotypes, and 47 GG genotypes. The results of statistical analysis between different genotypes and production traits are shown ...
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