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Method for assisting identification of hens laying green-shelled eggs and special primer pair thereof

A green-shell laying hen, an auxiliary identification technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, animal husbandry, etc., can solve the problems of prolonging the breeding cycle, high financial resources, human burden, and inaccurate gene positioning. Achieve the effects of shortening the breeding cycle, speeding up the breeding process and improving accuracy

Inactive Publication Date: 2011-01-05
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This requires a large number of test crosses, which not only prolongs the breeding cycle, but also causes a high financial and human burden
[0005] Although the mapping of the green shell gene has been carried out for more than 30 years, the gene has not yet been accurately located

Method used

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  • Method for assisting identification of hens laying green-shelled eggs and special primer pair thereof
  • Method for assisting identification of hens laying green-shelled eggs and special primer pair thereof
  • Method for assisting identification of hens laying green-shelled eggs and special primer pair thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1、13b

[0023] Example 1, Discovery of 13bp deletion polymorphism and design of primer pairs

[0024] Dongxiang green-shell layer hen is a famous green-shell layer breed in my country. Because the green-shell gene (O) is a dominant gene, there are still some heterozygotes carrying the recessive gene (o) in the population. The offspring produced between the homozygotes of the green-shell gene did not segregate in egg-laying traits, and all produced green-shell eggs. About 1:1 homozygous green shell gene and heterozygous green shell gene will be produced between the homozygous green shell gene and the heterozygous green shell gene, so that the non-green shell gene can be passed on. The offspring produced between the heterozygotes of the green-shell gene will produce individuals with brown-shell eggs (homozygous for the non-green-shell gene), resulting in an embarrassing situation of purebred being impure. That is, Dongxiang green-shell hens can be divided into three genotypes according...

Embodiment 2

[0032] Example 2, the application of primer pairs to assist in the identification of Dongxiang green-shell hens

[0033] 289 Dongxiang green-shell layer hens were purchased from Hualu Breeding Farm in Dongxiang County, Jiangxi Province. Cross each Dongxiang green-shell hen with the white Laihang chicken: if the offspring all produce green-shell eggs, the Dongxiang green-shell hen is homozygous for the green-shell gene; if the offspring have both green-shell egg-producing individuals and For individuals producing brown-shell eggs, the Dongxiang green-shell hens are heterozygous for the green-shell gene; if all offspring produce brown-shell eggs, the Dongxiang green-shell hens are homozygous for the non-green-shell gene. Among the 289 Dongxiang green-shell hens, 104 were homozygous for the non-green shell gene, 150 were homozygous for the green shell gene, and 35 were heterozygous for the green shell gene.

[0034] 1. Genomic DNA extraction

[0035] Blood was collected from th...

Embodiment 3

[0078] Embodiment 3, application primer pairs auxiliary identification green shell gene (O)

[0079] Given that the 13bp deletion is significantly associated with the green shell trait, and the marker falls within the finely mapped marker interval of the green shell gene, it is speculated that the marker (A) is likely to be related to the green shell gene (O) or has a close linkage relationship. By crossing Dongxiang green shell cock (OOAA) with Dongxiang brown shell hen (ooaa), F 1 Selfed, got a 152 F 2 A separate group of individuals. The linkage between the 13bpdel marker and the green shell gene was detected in the segregation population, and the results showed that the marker was tightly linked with the green shell gene (O), and the recombination rate was 0.04 (LOD=9.53). It can be seen that the probability of A and 0 being on the same haplotype is 96% (1-4%). Therefore, when the marker genotype was used to infer the green shell genotype, the accuracy could reach 92.16...

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Abstract

The invention discloses a method for assisting the identification of hens laying green-shelled eggs and a special primer pair thereof. The invention provides a primer pair formed by DNA (Deoxyribonucleic Acid) shown in a sequence (1) and a sequence (2) of a sequence table. The method of the invention comprises the following steps of: by using a genomic DNA as a template, carrying out PCR (Polymerase Chain Reaction) amplification with the primer pair; judging the hen to be tested as a candidate green eggshell gene homozygote if only one product is obtained from the PCR amplification and is as shown in a sequence (3); judging the hen to be tested as a candidate green eggshell gene heterozygote if two products are obtained from the PCR amplification and are respectively as shown in the sequence (3) and a sequence (4); and judging the hen to be tested as a candidate non-green eggshell gene homozygote if only one product is obtained from the PCR amplification and is as shown in the sequence (4). The invention finds a deletion marker of 13 bp significantly associated with the green eggshell property in a location area of the green eggshell gene so that both hens and cocks can be selected from the current chicken. The invention can overcome the defects of a conventional breeding method, shorten the breeding period and improve the breeding accuracy.

Description

technical field [0001] The invention relates to a method for assisting identification of green-shell laying hens and a pair of special primers. Background technique [0002] The eggshell color of poultry eggs can be divided into three types according to the main color system, white shell, brown shell and green shell (Lang, M.R., and J.W.Wells.1987.A review of eggshell pigmentation.World's Poult.Sci.J.43 : 238-245.). With its unique eggshell color, green-eggs have always attracted the attention of researchers at home and abroad. The more foreign research is the Chilean Araucano (Araucano) green-shell hens. In my country, the breeds that produce green-shell eggs include Sichuan Jiuyuan chicken, Henan Lushi chicken, Hubei Jianghan chicken and Jiangxi Dongxiang green-shell chicken. Among these breeds, Dongxiang green-shell hens are the most studied. [0003] Punnett (1933) found that the green shell trait is a quality trait controlled by a dominant single gene (Punnett RC. G...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68A01K67/027
Inventor 邓学梅王哲鹏王晓通吴常信
Owner CHINA AGRI UNIV
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