Chicken FABP1 gene molecular genetic marker related to chicken good production traits and application thereof
A technology of molecular genetic markers and production traits, applied to the application field of broiler chicken breeds
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Embodiment 1
[0026] Embodiment 1: the cloning of different breed chicken FABP1 gene partial DNA sequence
[0027] 1. Collection and processing of chicken blood: randomly selected disease-free healthy chickens, including 670 Luqin No. 3 chickens and 60 Langya chickens, blood was collected from the underwing vein, and EDTA was used for anticoagulant treatment.
[0028]2. Genomic DNA extraction: DNA was extracted using Tiangen blood DNA extraction kit. Use 200 μL of fresh, frozen or blood with various anticoagulants, add 20 μL proteinase K solution, mix well; then add 200 μL buffer GB, mix well and place at 70°C for 10 minutes; add 200 μL absolute ethanol, shake and mix well At this time, flocculent precipitates will appear; transfer all the obtained solution and flocculent precipitates to the adsorption column CB3 in the collection tube, centrifuge at 12000rpm for 30 seconds and discard the waste liquid; add 500μL buffer GD, centrifuge at 12000rpm for 30 seconds and Discard the waste soluti...
Embodiment 2
[0038] Embodiment 2: Use MboI enzyme to carry out specific enzyme digestion to the PCR product in embodiment 1
[0039] 1. Use MboI enzyme to specifically digest the target gene. The sites of action of these specific enzymes are:
[0040] 5'... ↓ GATC…3’
[0041] 3’…CTAG ↑ …5’
[0042] 2. See Table 3 for the enzyme digestion reaction system.
[0043] Table 3 MboI digestion reaction system
[0044]
[0045] After mixing the solutions of the reaction system according to the above system, place them in a constant temperature water bath at 37°C to react for 3-5 hours, so that the enzyme cleavage reaction can fully proceed, and then carry out subsequent tests.
[0046] 3. The digested products were analyzed by agarose gel electrophoresis. Then classify and summarize all samples according to different enzyme digestion results, and conclude different types.
[0047] There are three kinds of enzyme digestion results, namely complete mutation of SNP site, heterozygous mutati...
Embodiment 3
[0050] Example 3: Diagnostic application of molecular genetic markers prepared by the present invention in polymorphisms in different chicken populations
[0051] 1. Diagnosis in population polymorphism: 670 Luqin No. 3 chickens and 60 Langya chicken FABP1 genes were detected by specific enzyme digestion using the above-mentioned SNP polymorphism detection method.
[0052] 2. Frequency statistical analysis of SNP loci:
[0053] Genotype frequency refers to the ratio of the number of individuals of a certain genotype to the total number of individuals in a population. Paa=Aaa / N, where Paa represents the AA genotype frequency at a certain locus, Aaa represents the number of individuals with the AA genotype in the population, and N is the total number of the detection population.
[0054] Gene frequency refers to the relative ratio of the number of a certain gene to the total number of alleles in a population. The calculation formula can be written as: PA=(2NM+NA al +NA a2 +....
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