Molecular marker correlated to characteristics of maroon feathers of quails and application thereof
A molecular marker, quail technology, applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of lack of identification of homozygous genotypes and heterozygous genotypes in feather color, and save breeding costs. , The effect of short time and easy operation
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Embodiment 1
[0022] Example 1 Obtaining of Quail CCDC171 Gene Fragment and Establishment of Mutation Detection Method
[0023] 1. Extraction of quail genomic DNA
[0024] The experimental quail of the present invention is "Shendan No. 1" egg quail with yellow feather and chestnut feather male quail (the sample comes from the breeding quail farm of Hubei Shendan Health Food Co., Ltd.). Quail genomic DNA was extracted using a blood genomic DNA extraction kit (operated in accordance with the kit instructions) produced by Beijing Quanshijin Biotechnology Co., Ltd. The specific steps are as follows:
[0025] (1) Take 10 μL of quail anticoagulated blood, add 20 μL of proteinase K, 500 μL of BB3, vortex for 15 s to fully mix the sample, and incubate at room temperature for 10 min;
[0026] (2) Briefly centrifuge, transfer the entire solution into a spin column, centrifuge at 12,000×g for 1 min, and discard the filtrate;
[0027] (3) Add 500 μL solution CB3, centrifuge at 12000×g for 30 s, and d...
Embodiment 2
[0052] Example 2 The application of the molecular marker prepared by the present invention in the selection of sorrel feather heterozygous male quail
[0053] Using the molecular markers prepared by the present invention, the applicant detected the distribution frequencies of the two alleles of the quail CCDC171 gene in two populations of male quails with yellow feathers and sorrel feathers. The detection results are shown in Table 1.
[0054] Table 1 Distribution of alleles at mutation site 210 of CCDC171 gene in different plumage color male quail populations
[0055]
[0056] From the results in Table 1, it can be seen that the frequency of the A allele is 100% in male yellow-feathered quails, indicating that individuals with the AA genotype are all yellow-feathered and homozygous; The frequency of the allele is 90%, and the frequency of the A allele is 10%, indicating that in the sorrel male quail population, some individuals have a heterozygous AG genotype, and most ind...
Embodiment 3
[0057] Example 3 Application of the molecular markers prepared by the present invention in quail self-sexing mating lines
[0058] From the results in Example 2, it can be seen that the genotypes of male sorrel plumage quail are GG and GA, and the GA type is heterozygous. In this example, the above-mentioned genotypes are heterozygous individuals by the method of molecular marker-assisted selection. Selected to form a new group of sorrel feather male quail, breed offspring by crossing with yellow feather female quail, and observe the feather color and sex of the offspring.
[0059] After molecular marker-assisted selection, 11 heterozygous individuals whose sorrel feather genotype was GA were selected, and 132 yellow feather female quails were selected to form 11 families (each family had 1 sorrel feather male quail and 12 yellow feather quails). feather female quail) to breed offspring, the specific process is as follows:
[0060] 1. Breeding and egg collection
[0061] The...
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