Identification method of sex-linkage Luhua chicken genotype of Chinese native chickens

A chicken breed and Chinese technology, which is applied in the identification field of the sex-linked Luhua chicken genotype of native chicken breeds in my country, can solve the problems of unclear function of CDKN2A gene, undetermined specific mutation sites of phenotype, and deletion, etc., and achieves good application. Value, high accuracy, simple operation effect

Active Publication Date: 2019-07-16
SOUTH CHINA AGRI UNIV
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although it has been found that the CDKN2A gene is related to the sex-linked reed flower phenotype, the specific mutation site that affects its phenotype has not been determined, and the role of each SNP on the CDKN2A gene is also unclear, and the formation mechanism of the reed flower phenotype is still ongoing. exploring
The absence of genotypes and identification methods of sex-linked reed chickens in my country's native chicken breeds has seriously affected the breeding of breeders in my country

Method used

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  • Identification method of sex-linkage Luhua chicken genotype of Chinese native chickens
  • Identification method of sex-linkage Luhua chicken genotype of Chinese native chickens
  • Identification method of sex-linkage Luhua chicken genotype of Chinese native chickens

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Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1 A kind of method of detecting chicken sex-linked reed flower phenotype

[0038] SNP1 is the CDKN2A gene promoter region and 267bp upstream of the transcription initiation site. Primers were designed according to its position on the genome, and primers with high amplification specificity were screened out, and the amplification system and reaction program were optimized.

[0039] 1. Experimental method

[0040] Collect chicken blood samples, extract blood sample DNA, and amplify the blood sample DNA. The amplification primers are:

[0041] SNP1 upstream primer: 5'-GTTACCCGTTTCTGCTCCTTGACGC-3' (SEQ ID NO: 1);

[0042] SNP1 downstream primer: 5'-GCCCCCACCGTAAGAAAGCTCTCTC-3' (SEQ ID NO: 2).

[0043] The 50μL amplification system is as follows:

[0044]

[0045] SNP1 amplification program: pre-denaturation at 94°C for 2 minutes; denaturation at 98°C for 10 seconds, annealing at 60°C for 20 seconds, extension at 68°C for 50 seconds, a total of 40 cycles; ex...

Embodiment 2

[0049] Embodiment 2 A kind of method of detecting chicken sex-linked reed flower phenotype

[0050] SNP2 is the 380bp of intron 1 of CDKN2A gene. According to its position on the genome, primers were designed, and primers with high amplification specificity were screened out, and the amplification system and reaction program were optimized.

[0051] 1. Experimental method

[0052] Collect chicken blood samples, extract blood sample DNA, and amplify the blood sample DNA. The amplification primers are:

[0053] SNP2 upstream primer: 5'-GGTCGGCAGGCTATCTCTAT-3' (SEQ ID NO: 3);

[0054] SNP2 downstream primer: 5'-AGGTCCACGTATTCTCATCA-3' (SEQ ID NO: 4).

[0055] The 50μL amplification system is as follows:

[0056]

[0057] SNP2 amplification program: pre-denaturation at 94°C for 3 minutes; denaturation at 94°C for 30 seconds, annealing at 58°C for 30 seconds, extension at 72°C for 30 seconds, a total of 34 cycles; extension at 72°C for 5 minutes.

[0058] After purification,...

Embodiment 3

[0061] Embodiment 3 A kind of method of detecting chicken sex-linked reed flower phenotype

[0062] SNP3 is at 170bp of exon 1 of CDKN2A gene, and primers were designed according to its position on the genome, and primers with high amplification specificity were screened out, and the amplification system and reaction program were optimized.

[0063] 1. Experimental method

[0064] Collect chicken blood samples, extract blood sample DNA, and amplify the blood sample DNA. The amplification primers are:

[0065] SNP3 upstream primer: 5'-CTCTCCTGTTCCCATGACCTCTCG-3' (SEQ ID NO: 5);

[0066] SNP3 downstream primer: 5'-GATAGAGATAGCCTGCCGACCACC-3'(SEQ ID NO:6),

[0067] The 40μL amplification system is as follows:

[0068] Template DNA 1.5 μL

[0069] 10pmol primers 1.8μL each

[0070] 1.1×T3 Super PCR Mix 35μL

[0071] SNP3 amplification program: pre-denaturation at 98°C for 3 minutes; denaturation at 98°C for 10 seconds, annealing at 65°C for 10 seconds, extension at 72°C for ...

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Abstract

The invention discloses an interlocking SNP site combination related to the sex-linkage Luhua phenotype of Chinese native chickens. The SNP sites comprise SNP1, SNP2 and SNP3, wherein the SNP1 is a position of 267 bp of a CDKN2A gene promoter region and transcription initiation site upstream, the SNP2 is a position of 380 bp of CDKN2A gene intron 1, and the SNP3 is a position of 170 bp of CDKN2A gene exon 1. The invention finds the three SNP sites related to the sex-linkage Luhua chicken phenotype of Chinese native chickens for the first time, and establishes a method for detecting the phenotype and the genotype of sex-linkage Luhua chickens by the SNP sites. The method has the advantages of simple operation, high accuracy, good repeatability and time saving, is available to mutual detection, realizes 100% accuracy, can be used for cultivating auto-sexing commercial strains, performs auto-sexing by using feather colors, is high in accuracy and simple and convenient to operate, saves time and cost, has good application value, and is worthy of great popularization.

Description

technical field [0001] The invention relates to the technical field of molecular breeding of poultry, and more specifically relates to a method for identifying the genotype of sex-linked reed chicken of native chicken breeds in my country. Background technique [0002] As an important selection index in chicken breeding, feather color not only has certain ornamental value, but also has certain economic benefits. There are many kinds of feather colors of poultry, which can be divided into two types through biochemical and histological analysis: one is colored feathers, the other is colorless (white) feathers, and the difference of various colored feathers lies in the amount of melanin contained different, and affected by the migration and distribution of melanin. Further research found that the source of feather color is the result of the interaction between the pigment source gene CC and the oxidase gene OO, black is the final product of the chromogen oxidation reaction, re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 符蓉张细权郑茗张梓豪莫国东赵蒙蒙
Owner SOUTH CHINA AGRI UNIV
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