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A method and application for detecting five-toed traits of Beijing chicken

A technology for oil chicken and traits, which is applied in the field of detection of the five-toed traits of Beijing oil chicken, can solve the problems of easily affected by experimental conditions, complicated steps, low sensitivity, etc., and achieve accurate and reliable experimental results, high sensitivity and high degree of automation. Effect

Active Publication Date: 2020-02-21
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide a method that can accurately and efficiently detect the defect that the existing PCR-RFLP method is used to detect the existence of the five-toed traits of Beijing oil chicken, which is cumbersome, time-consuming, low-sensitivity, and detection results are easily affected by experimental conditions. Fluorescent Quantitative PCR Method for Detecting Five-toed Traits of Beijing Youji Chicken

Method used

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  • A method and application for detecting five-toed traits of Beijing chicken
  • A method and application for detecting five-toed traits of Beijing chicken
  • A method and application for detecting five-toed traits of Beijing chicken

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Embodiment 1

[0029] Example 1 Detection of mutation site rs80659072

[0030] Refer to the mutation site rs80659072 (NCBI accession number) and its surrounding sequence (nucleotide sequence as shown in SEQ ID NO: 5), design primers, upstream primer: 5'-ACATACCAAGAATGTGCATGTGC-3', downstream primer: 5'-TTTGAGGTAACTTCCTTGCTTAA -3', the length of the amplified product is 448bp.

[0031] Individuals of various toe shapes were randomly selected from a large flock of Beijing oil chickens, accumulatively 155. Anticoagulant blood was collected, genomic DNA was extracted using a DNA extraction kit (Tiangen Biochemical, Beijing), and the site and its surrounding sequences were subjected to PCR amplification using the above primers. After the amplified product was detected by 2% agarose gel, it was sequenced by Beijing Qingke Jiamei Biotechnology Co., Ltd. and reverse sequenced.

[0032] figure 1 The results of agarose gel electrophoresis detection after PCR amplification of some individuals mentio...

Embodiment 2

[0036] The establishment of embodiment 2 fluorescence quantitative PCR detection method

[0037] After repeated comparison, screening and verification, the present invention obtains a pair of chicken rs80659072 mutations according to the sequence of the ZRS region in the 5th intron of the LMBR1 gene and its vicinity (the nucleotide sequence is shown in SEQ ID NO: 5). Fluorescent quantitative PCR primers for the sites and TaqMan probes used in conjunction with the primers.

[0038] Upstream primer F: 5'-TCAGTGGCAAAAAACGAGCAAAAAT-3' (SEQ ID NO.1)

[0039]Downstream primer R: 5'-CACACAGAAATGAGTAGGAAGTCCAA-3' (SEQ ID NO.2).

[0040] The upstream primer consists of 25 bases, corresponding to base positions 8467207-8467231 of chicken No. 2 chromosome (www.ensembl.org, galgal4), and the downstream primer consists of 26 bases, corresponding to base positions 8467272-8467299 of chicken No. 2 chromosome (www.ensembl.org, galgal4), the length of the amplified fragment of the primer pai...

Embodiment 3

[0057] Embodiment 3 Different toe number phenotypes and genotype chicken genetic research

[0058] Males and hens with different toe number phenotypes and different genotype combinations were selected for mating, and the genetic rules of different genotypes were studied. The specific experimental operation is as follows:

[0059] (1) Double four-toed phenotype: 7 GG roosters, mated with 20 GG hens; 3 TT roosters, 7 TT hens, 7 GT hens; (2) Double five-toed phenotype : 8 TT cocks, with 20TT hens and 6 GT hens; 2 GT cocks and 2 GT hens.

[0060] Observation and registration of toe type were carried out on the offspring of the above mating types respectively. The results are shown in Table 2:

[0061] Table 2 Toe type results of chicken mating offspring with different toe number phenotypes and genotypes

[0062]

[0063] It can be seen from Table 2 that 100% of the offspring of CC type and CC type are double four-toed. The test results fully demonstrate that if the GG genot...

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Abstract

The invention discloses a method for detecting five-finger characters of Beijing fatty chickens. The method comprises the step of detecting the genotype of an rs80659072 mutation site in a ZRS area in a fifth intron of an LMBR1 gene by adopting a real-time fluorescent quantitative PCR (polymerase chain reaction) technique. The real-time fluorescent quantitative TaqMan SNP typing technique adopted by the method has the advantages of high detection speed, high sensitivity, high degree of automation and simplicity and convenience in operation, the type can be judged by only one PCR, and the whole reaction process is implemented in a closed tube, so that cross contamination is reduced. On the basis of the method, molecular markers are used for assisting breeding, so that a pure five-finger strain of the Beijing fatty chickens can be quickly constructed.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a method and application for detecting the traits of five toes of Beijing oil chicken. Background technique [0002] Beijing oil chicken is a well-known local breed in my country. It is native to the Beijing area. This breed of chicken has excellent meat and egg quality and is very popular among farmers and consumers. In recent years, it has been promoted in Beijing and most provinces and cities across the country, and the breeding volume and breeding scale have increased year by year. The chicken has a unique appearance and has the characteristics of five toes and three hairs (crested head, beard, and hairy legs). [0003] Among them, the five toes are the most obvious identification traits after slaughtering and marketing of Beijing Youji, which can be used as an important breed identification of Beijing Youji to help consumers make consumption choices. Therefore, it is of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/124C12Q2600/156C12Q2531/113C12Q2561/101
Inventor 初芹张剑张尧晏志勋王海宏耿爱莲刘华贵
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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