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Molecular marker for differentiating between simulated native eggs and native eggs and application

A technology of molecular markers and native eggs, applied in the field of molecular biology, achieves the effects of simple operation, accurate identification, and easy promotion and use at the grassroots level

Active Publication Date: 2019-03-08
JIANGSU INST OF POULTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there has been no research report on the identification of imitation native eggs and native eggs by molecular markers

Method used

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  • Molecular marker for differentiating between simulated native eggs and native eggs and application
  • Molecular marker for differentiating between simulated native eggs and native eggs and application
  • Molecular marker for differentiating between simulated native eggs and native eggs and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1 Establishment of molecular detection methods for imitation native eggs and native eggs

[0036] 1.1 Primer design

[0037] The female line of high-yielding powder-shell hens is derived from high-yielding layer hens, and has a different maternal origin from the female line of native chickens. The applicant conducted full-length sequencing of the mitochondrial genomes of the high-yielding layer population and the native chicken population, and found that the two populations had a variation at 1213bp , design primers to amplify the candidate site.

[0038] Forward primer (SEQ ID NO.1): 5'-CTGGACTACACCTGCGTTGCG-3'

[0039] Reverse primer (SEQ ID NO.2): 5'-TGCCTGATACCTGCTCCTTTT-3'

[0040] 1.2 PCR amplification

[0041] The PCR reaction system is: 10 μL of 2×PCR Mix, 0.5 μL of each 10 μmol / L forward primer, 1 μL of 50-100 μg / ml template DNA, and 8 μL of sterilized water.

[0042] The PCR reaction program was: 94°C for 5 minutes, 35 cycles (95°C for 30s, 55°C for 30s, 7...

Embodiment 2

[0046] Example 2 Application of the molecular marker of the present invention in native eggs and imitation native eggs

[0047] Collect the eggs of domestic chicken breeds (white-eared yellow chicken and Xianju chicken), high-yield small pink-shell egg breeds (Huafeng chicken and dwarf high-yield pink-shell chicken) eggs, 60 eggs for each variety, and a total of 240 eggs . The extraction of egg yolk membrane DNA refers to (Zhang Ying et al., Extraction and Application of Egg Yolk Membrane DNA, Research Progress in Animal Genetics and Breeding in China—Proceedings of the Fifteenth National Symposium on Animal Genetics and Breeding. 2009, 475).

[0048] Table 1 is the genotype distribution of 240 eggs of 4 varieties detected by the molecular marker of the present invention.

[0049]

[0050]As can be seen from Table 1, the eggs containing the CC type are all native eggs, and the eggs containing the TT genotype are all imitation earth eggs, that is, there are differences betw...

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Abstract

The invention discloses a molecular marker for differentiating between simulated native eggs and native eggs and an application. The molecular marker is of C / T alkaline mutation at the 1213bp of a mitochondria genome. The gene sequence of the native egg of the molecular marker is C while that of the simulated native egg is T. By amplifying the gene segment containing the molecules by means of theprimers SEQ ID NO. 1 and SEQ ID NO.2 and performing digestion on the amplified product with a restriction enzyme BspMI, the genotype of a mutant site is judged according to a digestion result. The method is simple to operate and reliable in structure. The simulated native eggs and the native eggs can be differentiated quickly and accurately according to the method. The method is easy to operate, low in cost and convenient to popularize and apply in a basic level, and has a wide market application prospect.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular marker for identifying imitation native eggs and native eggs and its application. Background technique [0002] Compared with brown-shell and white-shell eggs, Chinese citizens prefer pink eggs, because the eggshell color of local breeds in my country is mostly pink, and people generally call eggs produced by local breeds native eggs. In order to meet the needs of consumers, most of the powder-shell eggs in the market now are eggs produced by the high-yield powder-shell layer breeds that are generally hybrids of foreign brown-shell breeds and white-shell breeds. We call them imitation soil eggs. [0003] The high-yield powder-shell breeds can lay more than 300 eggs per year, while the local native chickens only have about 120 or less. Although the appearance of imitation soil eggs and soil eggs is similar, the quality is very different. It is di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 高玉时贾晓旭陆俊贤唐修君樊艳凤姬改革葛庆联黄胜海唐梦君刘茵茵张静马丽娜陈大伟张小燕周倩马尹鹏
Owner JIANGSU INST OF POULTRY SCI