Genetic mark using SNP of third exon of CATSPER4 gene as sperm quality property of pigs
A technology of genetic markers and quality traits, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problem of low heritability of reproductive traits in boars
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Embodiment 1
[0019] Example 1: Obtaining of porcine CATSPER4 gene fragment and establishment of polymorphism detection method
[0020] Log in to the Ensembl database to download the porcine CATSPER4 gene sequence (the accession number is ENSSSCG00000026526), find the screened SNP site in the sequence, and use Primer Premier 5 software to design primers. The sequence of the forward primer is shown in SEQ ID NO: 2 in the sequence table, that is, CATSPER4-AlwNI-F: GCA ATG ACC ACAGCG AAG A, and the sequence of the reverse primer is shown in SEQ ID NO: 3 in the sequence table, that is, CATSPER4-AlwNI -R: TGC TCAGGC CCG TTA GAT C. Amplified to obtain a 410bp target fragment (the sequence of this fragment is shown in SEQ ID NO: 1 (see figure 2 ). The PCR reaction system is 25 μl, including 1 μl template DNA, 0.5 μl each primer, 12.5 μl PCR Mix, and finally add 10.5 μl deionized water to a total volume of 25 μl; PCR reaction program: 94 ° C pre-denaturation for 5 minutes; then Denaturation a...
Embodiment 2
[0022] Example 2: Polymorphic distribution of genetic markers prepared by the present invention in different pig herds
[0023] The extraction of pig genomic DNA (sample is shown in Table 1) was carried out with reference to the method reported by Xiong Yuan, "Introduction to Pig Biochemistry and Molecular Genetic Experiments", China Agricultural Press, 1999.
[0024] The PCR-AlwNI-RFLP polymorphism of porcine CATSPER4 gene was detected in three foreign blood-related pig populations, and the detection results are shown in Table 1.
[0025] Table 1 Distribution results of porcine CATSPER4 gene PCR-AlwNI-RFLP in different pig breeds
[0026]
[0027] Note: The above group materials are all species publicly promoted in China.
[0028] The results showed that there were 3 genotypes in Duroc, Large White and Landrace pigs. In the Duroc pig population, the G allele frequency is 0.83, which is the dominant allele; in the Large White pig population, the G allele frequency and T all...
Embodiment 3
[0029] Example 3: Association analysis of genetic markers prepared by the present invention and semen quality traits
[0030] In order to determine whether the pig CATSPER4 gene PCR-AlwNI-RFLP is related to the difference in the quality of pig semen, the Duroc pigs, Large White pigs and Landrace pigs bred in the breeding farm of Guangxi Yangxiang Co., Ltd. were selected as test materials, and the pigs established in Example 1 were used. AlwNI-RFLP method was used to detect polymorphisms, and the correlation between different genotypes of pig AlwNI-RFLP and pig semen quality traits was analyzed. The SAS statistical software (SAS Institute Inc, Version 9.4) 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:
[0031] Y ij =μ+G i +F j +e ij
[0032] Y ij is the trait phenotype value, μ is the average value, G i ...
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