A genetic marker related to pig semen quality traits and its application
A technology for genetic markers and quality traits, applied in the field of swine genetic marker screening, can solve the problems of slow progress and low boar heritability
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Embodiment 1
[0025] Example 1: The acquisition of pig RNF4 gene fragments and the establishment of polymorphism detection methods
[0026] Log in to the NCBI database, download the porcine RNF4 gene sequence as the target sequence, and use Primer Premier 5 software to design primers. The sequence of the primer is shown in the sequence table SEQ ID NO: 1 and SEQ ID NO: 2, and the specific sequence is as follows: forward primer RNF4-F: AGA ACG CTA ACA CTT GCC, reverse primer RNF4-R: CAC GGT GAC CTC TGA CTC. The amplified region includes the 649bp genomic nucleotide sequence of the 3' untranslated region of the pig RNF4 gene, such as figure 1shown. The PCR reaction system was 25 μl, in which the template DNA was 50 ng, the concentration of dNTPs was 200 μmol / L, the concentration of each primer was 0.4 μmol / L, 3 U of Taq DNA polymerase (Biostar International, Canada), and deionized water was added to a total volume of 25 μl ;PCR reaction program: pre-denaturation at 94°C for 4min; then den...
Embodiment 2
[0028] Example 2: Detection of polymorphism distribution of genetic markers screened by the present invention in different pig herds
[0029] The extraction of pig genome DNA (sample is shown in Table 1) is carried out with reference to the method reported in 1999 in Xiong Yuan's "Introduction to Pig Biochemistry and Molecular Genetic Experiments" China Agricultural Press.
[0030] The PCR-AluI-RFLP polymorphism of the porcine RNF4 gene was detected in three foreign blood-related pig populations, and the detection results are shown in Table 1. The results showed that there were three genotypes in Duroc, Large White and Landrace pigs; in the Duroc population, the C allele frequency was 0.84, which was the dominant allele; in the Large White pig population, the T allele The frequency is 0.78, which is the dominant allele; in the Landrace pig population, the C allele frequency and T allele frequency are 0.44 and 0.56, respectively, which are alleles with no obvious advantages (se...
Embodiment 3
[0034] Example 3: Application of the association analysis between the genetic markers screened by the present invention and pig semen quality traits
[0035] In order to determine whether the pig RNF4 gene PCR-AluI-RFLP is related to the difference in the quality of pig semen, the Duroc pigs, Large White pigs and Landrace pigs bred in Guangxi Yangxiang Pig Farm were selected as test materials, and the AluI-RFLP established in Example 1 was used. Methods The polymorphism was detected, and the correlation between different genotypes of porcine AluI-RFLP and porcine semen quality traits was analyzed. The SAS statistical software (SAS Institute Inc, Version 8.0) GLM program was used for single-marker analysis of variance, and the REG program was used to calculate the additive effect and dominant effect of genes, and the significance test was carried out. The model used was:
[0036] Y ij =μ+G i +F j +e ij
[0037] Y ij is the trait phenotype value, μ is the average value, G ...
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