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Molecular diagnosis of sex-linked dwarf chicken and its uses in dwarf chicken breeding

A small chicken and mutant technology, applied in the application field of dwarf chicken breeding, can solve the problems of the uniformity of the offspring, the lack of systematic establishment of detection, and the irregular production performance of the offspring, etc.

Inactive Publication Date: 2007-07-11
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] It has been proved that the sex-linked dwarfism of chickens is caused by the mutation of its growth hormone receptor gene GHR (its coding sequence is shown in SEQ ID NO.7 in the sequence table), and the sites of GHR gene mutations of different dwarf chickens are not the same. Not the same, it has been found that there are at least 4 different mutations that can lead to the production of dwarf chickens. The binding activity of GH and GHR in dwarf chickens caused by different reasons is different, and thus it will cause dwarf chickens to have different body weight and shin length. Inconsistent performance on
If different individuals in a breed are short due to different reasons, it will seriously affect the uniformity of their body weight and shank length. When using it for crossbreeding, the production performance of their offspring is also extremely irregular, which makes it difficult to use dwarf chickens When breeding, the uniformity of offspring is affected to a certain extent
Different dwarf chicken flocks are caused by different mutations, and some dwarf chicken flocks have multiple sex-linked mutations. At present, no method for detecting these mutations has been systematically established, and the effects of different mutations on body weight and shank length have not been studied. Influence and its interaction

Method used

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  • Molecular diagnosis of sex-linked dwarf chicken and its uses in dwarf chicken breeding

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The detection of embodiment 1 type II mutation:

[0036] Primers (reference sequence GeneBank accession number: AC187590):

[0037] 5'-GGCAGAAACCCCAAGTTATG-3'

[0038] 5'-TCGGGACAGATCAAAGACAAT-3'

[0039] PCR amplification conditions:

[0040] Due to the longer fragments, the PCR amplification conditions are different, and the denaturation, annealing, and extension times are all longer. The PCR reaction program: 94°C for 3min, 35 cycles (94°C for 45s, 56°C for 45s, 72°C for 2min), Extend for 10 min and store at 4°C. Table 2 is the reaction system of PCR.

[0041] Table 2 PCR reaction mixture system

[0042] Template DNA (2.5ng / μL)

[0043] The amplified product was electrophoresed on 1.0% agarose gel, and the size of the fragment formed after deletion was 253bp, while the size of the fragment without deletion was 2026bp, so the genotype can be directly judged according to the size of the product fragment. The test results are shown in Figure 1. Normal ch...

Embodiment 2

[0044] The detection of embodiment 2 I and type IV mutation:

[0045] Primer C: 5'-ACACATCCTACACCTCGATTTGG-3'

[0046] 5'-CGAAAGCTTACAGAATCACAC-3'

[0047] PCR reaction program: 94°C for 3min, 35 cycles (94°C for 30s, 58°C for 30s, 72°C for 30s), extension for 7min, and storage at 4°C. Table 3 is the reaction system of PCR.

[0048] Table 3 PCR reaction mixture system

[0049] ExTaq (5U / μL)

10×Ex Taq Buffer

dNTP Mixture (2.5mM each)

Template DNA (2.5ng / μL)

Upper Primer (25μM)

Lower Primer (25μM)

Sterile distilled water

0.125μL

2.5μL

2.5μL

1μL

0.5μL

0.5μL

Up to 25μL

[0050] Take 5 μL of PCR amplification product mixed with 2 μL of bromophenol blue, and use DNAMarker as a reference, the concentration of agarose gel is 1.0% (containing 0.5ug / mL EB dye), 1 × TAE as a buffer, The electrophoresis is about 45min, and the voltage is 4-5V / cm. After the electrophoresis is over, observe whether there is an amp...

Embodiment 3

[0063] The detection of embodiment 3 type III mutations:

[0064] Primer 5'-CCCAGGCTCTCAACAGTG-3'

[0065] 5'-TGAGTAAAGGAAACATAGAGG-3'

[0066] Amplification conditions, reaction system and genotype judgment are the same as the detection of I and IV mutations.

[0067] The detection results are shown in Figure 3, only one genotype appeared, and no mutation was found after being sent for sequencing, indicating that there was no mutation at this site in each chicken breed.

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Abstract

The invention discloses a molecular diagonizing method of sex chained dwarf chick and application to breed dwarf chick, which is characterized by the following: judging the caused reason from four mutation points of growth hormone acceptor; obtaining the mutation number; purifying the dwarf chicken; improving weight and tibia length and evenness.

Description

technical field [0001] The invention relates to a molecular marker assisted breeding method, in particular to the molecular diagnosis of sex-linked dwarf chickens and its application in dwarf chicken breeding. Background technique [0002] The adult weight of dwarf chickens is only 60-70% of normal chickens, and the adult weight of heterozygous offspring mated with normal chickens is also 90% of normal chickens, but the egg production performance of dwarf chickens is not lower than that of normal chickens. Due to their small size, dwarf chickens require less nutrients to maintain them, and dwarf chickens have a higher feed conversion rate than their normal siblings, thus saving a lot of feed. And because of its small size, the area occupied by the house is also reduced by 40% compared with normal chickens. The number of breeding per unit area and the number of poultry products provided are large, which reduces the feeding cost and improves economic benefits. In the product...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张细权欧阳建华曾华
Owner SOUTH CHINA AGRI UNIV
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