Molecular marker, related to swine semen quality character, in tenth intron of swine SPATA6 gene and application
A molecular marker and quality trait technology, applied in DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc.
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Embodiment 1
[0021] Example 1 Obtaining of pig SPATA6 gene fragments and establishment of polymorphism detection method
[0022] 1. Extraction of pig genomic DNA
[0023] The test pig breeds of the present invention are Duroc pigs and Large White pigs, and the samples are from Huazhong Agricultural University. The extraction of pig genomic DNA adopts the genomic DNA kit (operated according to the instructions of the kit) produced by Beijing Biotech Biotechnology Co., Ltd., and the specific steps are as follows:
[0024] (1) Aspirate 100 μL sperm pellet; add 400 μL ddH 2 O. 100 μL of 5* sperm extract and 10 μL of proteinase K (20 mg / mL) were thoroughly mixed and bathed in a 55°C water bath until fully digested and transparent.
[0025] (2) Add 200 μL of binding solution CB, mix thoroughly by inversion, and place at 70°C for 10 minutes.
[0026] (3) After cooling, add 100 μL of isopropanol, vigorously invert and mix well.
[0027] (4) Use a 1 mL pipette tip to absorb the above mixture, a...
Embodiment 2
[0049] Example 2 Detection of Polymorphism Distribution of Molecular Genetic Markers Prepared in the Present Invention in Duroc Pigs and Large White Pigs
[0050] In this example, the polymorphisms of the tenth intron rs341061477 and rs331255092 sites of the porcine SPATA6 gene were detected in Duroc pigs and Large White pig populations respectively, and the detection results are shown in Table 1.
[0051] Table 1 - Genotype frequency and allele frequency of SPATA6 gene rs331255092 and rs341061477 in Duroc and Large White pigs
[0052]
[0053] The results showed that the allele A of rs341061477 locus was the dominant allele in Large White pigs, but the difference was not significant in Duroc pigs; the frequency of allele T in rs331255092 locus was 0.74 in Duroc pigs, which was the dominant allele , the frequency of allele C in Large White pigs is 0.76, which is the dominant allele (see Table 1).
Embodiment 3
[0054] Example 3 Association Analysis and Application of Molecular Genetic Markers Prepared in the Present Invention and Pig Litter Size Traits
[0055] In order to determine whether the rs341061477 and rs331255092 sites of the tenth intron of the pig SPATA6 gene are related to the difference in the quality of pig semen, the method established in Example 1 was used for polymorphism detection, and the relationship between the different genotypes of the pig SPATA6 gene and the quality of pig semen was analyzed. Correlation. The SAS statistical software (SAS Institute Inc, Version 9.1) 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:
[0056] Yij=μ+Gi+Fj+eij
[0057] Yij is the trait phenotype value, μ is the average value, Gi is the genotype effect (including gene additive effect and dominant effect; additive effe...
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