A molecular marker method for predicting and identifying chicken belly fat mass and its application

A belly fat, molecular marker technology, applied in the field of animal molecular genetics, to achieve the effect of low cost, high accuracy and accelerated breeding process

Active Publication Date: 2021-03-12
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no reports have been found on the function of chicken PCSK1 gene. At the same time, there is no method for selecting chicken abdominal fat content through PCSK1 gene molecular markers, and there is no SNP site c.1880C>T upstream and downstream 190bp Sequence design of primers and molecular markers for screening genotype reports

Method used

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  • A molecular marker method for predicting and identifying chicken belly fat mass and its application
  • A molecular marker method for predicting and identifying chicken belly fat mass and its application
  • A molecular marker method for predicting and identifying chicken belly fat mass and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 buffer solution preparation and primer design

[0029] 1. Experimental Animals and Determination of Traits

[0030] Broiler chickens bred by Northeast Agricultural University are high-fat and low-fat dual-selection lines with 542 roosters in the 19th generation and 685 roosters in the 20th generation; AA broiler random group of 348 chickens. Blood was collected from wing veins of high-fat and low-fat broilers and AA broilers at 7 weeks of age respectively, and EDTA-Na 2 Anticoagulation. The live weight was measured before slaughter at 7 weeks old, and the abdominal fat weight was measured after slaughter, and divided by the live weight at 7 weeks old to calculate the abdominal fat rate.

[0031] 2. Drugs and enzymes

[0032] Tris (Tris), Sigma Chemicals Co; Tris saturated phenol, Beijing Dingguo Biotechnology Development Center; Proteinase K (Proteinase K), MMERCK Co; DL 2000, Dalian Bao Biological Company; dNTP (dATP; dTTP; dCTP; dGTP), Taq enzyme, DNA...

Embodiment 2

[0047] Example 2 DNA extraction, product amplification and electrophoresis separation

[0048] 1. Extraction of chicken DNA

[0049] The following two methods can be used to extract a small sample of chicken DNA:

[0050] method one:

[0051] (1) Take 20 μl of anticoagulated blood, add 500 μl of poultry lysate, add proteinase K to a final concentration of 100-200 μg / ml, mix and digest at 55°C for 12 hours until there are no sticky lumps in the solution.

[0052] (2) Cool the solution to room temperature, add 5M NaCl to a final concentration of 1.5M, and mix for 10 minutes. Add an equal volume of phenol / chloroform and invert the centrifuge tube repeatedly for 10 min.

[0053] (3) Centrifuge at 12,000 rpm for 10 minutes at room temperature. Take the supernatant, add an equal volume of chloroform and mix for 10 min.

[0054] (4) Centrifuge at 12,000 rpm for 10 minutes at room temperature. Take 2 times the volume of supernatant to precipitate DNA with absolute ethanol.

[0...

Embodiment 3

[0080] Embodiment 3 Statistical Model Establishment

[0081] According to the characteristics of the group, construct the following linear model:

[0082] Y=μ+G+F+D(F)+BW7+e①

[0083] Y=μ+G+S+G×S+F+D(F)+BW7+e②

[0084] Y is the observed value of traits, μ is the group mean, G is the genotype fixed effect, S is the sex fixed effect, F is the random effect of the family, D(F) is the random effect of the hen within the family, BW7 is used as the covariance variable, e is the remaining value. Model ① was used to analyze the correlation between locus polymorphisms and abdominal fat and other traits in the 19th and 20th generation populations of high-fat and low-fat broiler chickens in Northeast Agricultural University; model ② was used to analyze locus in AA broiler random populations. Association of point polymorphisms with traits such as abdominal fat. The statistical software JMP 7.0 (SAS Institute, 2000) was used to test the correlation between population genotypes and trai...

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Abstract

The invention discloses a molecular marker method and application for predicting and identifying chicken abdominal fat mass, belonging to the technical field of animal molecular genetics. The method provided by the present invention is to design primers according to the SNP site c.1880C>T upstream and downstream 190bp sequence of the egg white precursor processing enzyme 1 gene, and then carry out PCR amplification of chicken genomic DNA according to the obtained primers, and then use restriction endonuclease Enzyme digests the amplified product, then electrophoresis separates the digested product, and finally uses PCR‑RFLP technology to analyze gene polymorphisms to obtain chicken abdominal fat marker genotypes. The method provided by the invention has the characteristics of simple operation, low cost and high precision, can carry out automatic detection, is an effective molecular marker breeding method, and can greatly speed up the breeding process of chickens.

Description

technical field [0001] The invention relates to a molecular marker method and application for predicting and identifying chicken abdominal fat mass, belonging to the technical field of animal molecular genetics. Background technique [0002] The broiler industry has a good development prospect in the world, especially in developing countries such as China. Over the past half century, remarkable progress has been made in breeding based on phenotypic value-based selection, resulting in significant increases in growth rate and meat production in broiler chickens. With the rapid development of my country's broiler industry, the physiological discomforts and related diseases of broilers have also increased, such as excessive body fat accumulation, ascites syndrome, sudden death syndrome, leg diseases, and decreased immune function of the body. The economic loss to broiler producers is obvious. Excessive deposition of broiler body fat (especially abdominal fat) will not only caus...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858C12Q1/6888
CPCC12Q1/6858C12Q1/6888C12Q2600/124C12Q2531/113C12Q2521/301C12Q2565/125
Inventor 张慧李辉杨莉莉
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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