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Method and primer combination for effectively identifying Luyuan chicken breeds

A combination of primers and deer identification technology, applied in the field of poultry genetic resources and biology, can solve problems affecting the accuracy of identification results

Pending Publication Date: 2021-04-09
JIANGSU INST OF POULTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Chinese invention patent CN1055950733B discloses an identification method for Luyuan chicken species. This method uses a set of specific SNPs sites on the Z chromosome of Luyuan chicken, and the physical positions of different sites are relatively close, which will affect the identification to a certain extent. Accuracy of results

Method used

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  • Method and primer combination for effectively identifying Luyuan chicken breeds
  • Method and primer combination for effectively identifying Luyuan chicken breeds
  • Method and primer combination for effectively identifying Luyuan chicken breeds

Examples

Experimental program
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Effect test

Embodiment 1

[0130] Example 1 Simplified genome sequencing screening of Luyuan chicken species-specific SNPs / INDELs markers

[0131] (one)

[0132] For sample: including Luyuan Chicken (20), Jining Hundred Day Chicken (20), Langya Chicken (20), Shouguang Chicken (20), Wenshang Reed Chicken (20), Rugao Yellow Chicken (20 ), Langshan Chicken (20), Taihu Chicken (20), Liyang Chicken (20), Xuhai Chicken (20), Longsheng Phoenix Chicken (20), Minqing Maojiao Chicken (20) , Huaibei No-Ma chickens (20), half male and half female for each species. 0.5-1.0ml of blood was collected from the wing vein, placed in an anticoagulant tube, and stored at -70°C.

[0133] Genomic DNA was extracted by CTAB method, digested with restriction endonucleases EcoRI and NIaIII, and then sequence adapters were added to construct a small fragment library for GBS simplified genome sequencing.

[0134] In order to check the efficiency of enzyme digestion and facilitate subsequent sequence analysis, electronic enzyme d...

Embodiment 2

[0187] Embodiment 2 A kind of method for effectively identifying Luyuan chicken

[0188] The method includes, using the following specific genetic loci for identification:

[0189] Luyuan_tag1 is located on chicken chromosome 1, at physical position 157669776, between gene MZT1 and gene LOC112532749, and the genotype of Luyuan chicken is TT;

[0190] Luyuan_tag2 is located on chicken chromosome 2, at physical location 148438251, the first intron of the gene RHPN1, and the genotype of Luyuan chicken is CC;

[0191] Luyuan_tag3 is located on chicken chromosome 3, at physical position 27832013, the first intron of gene LOC421419, the genotype of Luyuan chicken is TGTAGA / / TGTAGA, and the mutant genotype is the deletion of the TGTAGA segment;

[0192] Luyuan_tag4 is located on chicken chromosome 3, at physical position 38286930, the 10th intron of gene RBM34, and the genotype of Luyuan chicken is GG;

[0193] Luyuan_tag5 is located on chicken chromosome 4, at physical position 28...

Embodiment 3

[0201] Embodiment 3 A kind of method for effectively identifying Luyuan chicken

[0202] The method includes, using the following specific genetic loci for identification: Luyuan_tag1 is located on chicken chromosome 1, at physical position 157669776, between gene MZT1 and gene LOC112532749, and the genotype of Luyuan chicken is TT;

[0203] Luyuan_tag2 is located on chicken chromosome 2, at physical location 148438251, the first intron of the gene RHPN1, and the genotype of Luyuan chicken is CC;

[0204] Luyuan_tag3 is located on chicken chromosome 3, at physical position 27832013, the first intron of gene LOC421419, the genotype of Luyuan chicken is TGTAGA / / TGTAGA, and the mutant genotype is the deletion of the TGTAGA segment;

[0205] Luyuan_tag6 is located on chicken chromosome 4, at physical location 17182490, in the gene IL1RAPL2UTR3 region, and the genotype of Luyuan chicken is --mutant type is TT;

[0206] Luyuan_tag9 is located on chicken chromosome 11, at physical l...

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Abstract

The invention relates to the technical field of poultry genetic resources and biology, in particular to a method and primer combination for effectively identifying Luyuan chicken breeds. According to the invention, a group of Luyuan chicken specific genetic loci are obtained by screening and include SNPs loci and INDELs loci. The specific genetic loci are distributed on different chromosomes and are used as identification loci, so that the accuracy and flexibility of a detection test are improved. Secondly, the INDELs loci capable of being effectively used for identifying the Luyuan chicken breeds are obtained by screening for the first time. The identification rate of the genetic specific loci obtained by screening on Luyuan chickens is 80% or above. The identification method provided by the invention can more effectively realize the identification of the Luyuan chickens and is high in practicability; and the method, the primer combination and a kit can be widely popularized and applied, and have important significance for identification and protection of the Luyuan chickens.

Description

technical field [0001] The invention relates to the fields of poultry genetic resources and biotechnology, in particular to a method for effectively identifying Luyuan chicken species and a combination of primers. Background technique [0002] "Beggar Chicken" is well-known both at home and abroad, and its authentic method is to use Luyuan Chicken as an ingredient. Luyuan chicken, also known as Luyuan big chicken, is a dual-purpose chicken breed, named after it was produced in Luyuan Town, Shazhou County, Jiangsu Province. In recent years, thanks to the efforts of all walks of life, the brand of Luyuan Chicken has gained little influence. However, due to the low threshold of breeding technology, other local chicken breeds or hybrid Luyuan chickens are often used as purebred Luyuan chickens, which seriously affects the brand image of Luyuan chickens. Therefore, it is urgent to provide a method for effectively identifying Luyuan chicken species. [0003] Chinese invention p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/172
Inventor 王克华韩威曲亮李国辉窦套存王星果
Owner JIANGSU INST OF POULTRY SCI
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