Xupu goose fatty liver fat deposition A-FABP (adipocyte-fatty acid-binding protein) gene and genetic marking method
A technology of fat deposition and Xupu geese, applied in genetic engineering, plant genetic improvement, botany equipment and methods, etc., to achieve significant application value and economic benefits
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Embodiment 1
[0053] Example 1 SNP screening of genes related to fat deposition in fat liver of Xupu goose
[0054] Sixty healthy Xupu geese of the same age were selected for the filling-feeding experiment. After 28 days of slaughter, the livers of Xupu geese were weighed, and the genomic DNA was extracted from extreme samples (large and small livers).
[0055] According to the goose A-FABP gene sequence in GenBank, Primer5.0 was used to design primers covering the entire sequence of exon 2 to exon 3 of the gene. The primer sequence is: 5'-CCCAAATAAGATAAGAACAC-3' (forward), 5' -ACATCCACCAGACAAGAA-3' (reverse), synthesized by Hunan Qingke Biotechnology Co., Ltd. The genomic DNA of Xupu goose was amplified with the above primers respectively.
[0056] PCR: 20 μl reaction system: 12 μl of 2×Master mix, 0.8 μl each of upstream and downstream primers (10 μmol / L), 1 μl of DNA template, and make up to the required volume with ultrapure water.
[0057] PCR amplification program: pre-denaturation...
Embodiment 2
[0061] Example 2 Verification of SNPs related to fat deposition in Xupu goose fat liver in production practice
[0062] Twenty-one healthy Xupu geese aged 80 days and under the same feeding and management conditions were selected for the experiment. After 28 days of feeding, all the tested Xupu geese were slaughtered, and the weight of the liver was weighed.
[0063] Genomic DNA extracted from 21 Xupu geese pectoralis tissue samples was amplified using the primers in Example 1, and the SNPs were detected according to the method in Example 1.
[0064] 1. PCR
[0065] The PCR reaction system is shown in the table below:
[0066] Table 1 PCR reaction system
[0067]
[0068] The PCR cycling parameters are as follows:
[0069] Pre-denaturation at 95°C for 5min; denaturation at 95°C for 30s, annealing at 50.3°C for 30s, extension at 72°C for 1min, a total of 35 cycles; extension at 72°C for 10min, and finally storage at 4°C.
[0070] 2. Analysis of genotype frequency of A-F...
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