Piao chicken rumpless gene, and method, primer and kit for detecting rumpless trait of Piao Chicken
A gene detection and trait technology, applied in the field of mutation or genetic engineering, can solve the problems of indistinguishability of tailless chickens, difficulties in conservation and breeding of chickens, and unclear genetic rules and genetic basis of tailless chickens. High accuracy, time saving, low cost effect
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Embodiment 1
[0036] Example 1 Chicken hybridization test and its genetic law
[0037] Hybridization experiment 1: The hybridization experiment of Piaoji roosters and White Rock hens was carried out in Xiaosiao Progenitor Broiler Breeder Farm in Yunnan Province. The male parent of the cross-breeding combination comes from the Tailless Scooter Hen of Yunling Guangda Piao Chicken Original Breeding Conservation Farm in Zhenyuan, and the female parent is the White Rock hen of the Tego matching line.
[0038] Hybridization experiment 2: the hybridization experiment of Bailuo rooster and Piaoji hen, which was carried out in Yunling, Zhenyuan Guangda Piaoji Original Species Preservation Farm. The male parent of the hybrid combination is the Di Gao mating line White Rock rooster from the Xiaosiao Progenitor Broiler Breeder Farm in Yunnan Province, and the female parent is the koala hen.
[0039] Hybridization experiment 3: The hybridization experiment of Piaoji roosters and Hailan layer hens was c...
Embodiment 2
[0048] The acquisition of embodiment 2 chicken IRX1 gene polymorphic sites
[0049] 1 Chicken blood sample collection
[0050] In the sample collection, the sex, age, feather color, tail traits and pedigree information of each chicken were recorded in detail, and 2-3ml of whole blood was collected from the wing vein using a heparin sodium anticoagulated vacuum blood collection tube, transported to the laboratory at 4°C, and then Store at -20°C.
[0051] 2 Genomic DNA Extraction
[0052] Chicken wing vein blood collection, all blood samples were 1mL / bird, anticoagulated with EDTA, and the blood samples were frozen at -20°C. Adopt conventional phenol-chloroform method to extract genomic DNA from above-mentioned chicken blood, detect with 1.0% agarose electrophoresis, can see that DNA band is clear, bright, without miscellaneous band, fully meets the requirement of follow-up PCR test ( figure 2 ).
[0053] 3 Primer design and synthesis
[0054] According to the IRX1 gene co...
Embodiment 3I
[0070] Example 3 Association analysis of single SNP of IRX1 gene and tailless trait
[0071] The association analysis between a single SNP and chicken tail development was analyzed by one-way analysis of variance in SPSS. If P<0.05, it was significant.
[0072] Using the logistic regression method to analyze the correlation between single SNPs of the IRX1 gene and the tailless trait of the chicken under the codominant mode of inheritance, it was found that the SNP2 and SNP5 loci were significantly correlated with the tailless trait of the chicken (P0.05). IRX1 gene SNP3 site genotype G / G tailed group frequency (95.5%) is extremely significantly higher than the no-tailed group 0 (0%) frequency, suggesting that G / G type is a tailed genotype; IRX1 gene SNP8 site The frequency (95.5%) of the tailed group of genotype A / A is extremely significantly higher than the 0 (0%) frequency of the tailless group, suggesting that type A / A is a tailed genotype. The results are shown in Table 6....
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