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A molecular marker related to anti-parasitic infection traits of goats and its application

An anti-parasitic infection, molecular marker technology, used in applications, microbial determination/inspection, biochemical equipment and methods, etc.

Active Publication Date: 2016-09-07
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the study of goats, the research on the regulatory role of the TLR gene family in parasite infection has not yet appeared, but there are many studies on TLR regulation of parasite infection in other species, for example, the TLR2-mediated inflammatory response triggered by malaria infection is controlled It is considered to be a significant factor in inhibiting the growth of pathogens and disease control (Durai P et al., Structure and dynamic behavior of Toll-like receptor2subfamily triggered by malarial glycosylphosphatidylinositols of Plasmodium falciparum. FEBSJournal, 2013, 280(23): 6196-6212.)

Method used

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  • A molecular marker related to anti-parasitic infection traits of goats and its application
  • A molecular marker related to anti-parasitic infection traits of goats and its application
  • A molecular marker related to anti-parasitic infection traits of goats and its application

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Experimental program
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Effect test

Embodiment 1

[0035] (1) Partial DNA sequence amplification of TLR2 gene

[0036] (1) Extraction and purification of goat ear tissue-like DNA

[0037] All goat DNA samples were extracted using the TIANamp Genomic DNA Kit kit (operate according to the instructions of the kit), and the specific steps are as follows:

[0038] (1) Put the ear sample into a 2ml EP tube, wipe clean ophthalmic surgical scissors with alcohol cotton and cut into a paste, then add 200μl buffer GA.

[0039] (2) Add 20 μl proteinase K solution, mix well and place in a 56°C water bath for overnight digestion.

[0040] (3) Add 200 μl of buffer solution GB, mix thoroughly by inversion, place at 70°C for 10 minutes, the solution should become clear, and briefly centrifuge to remove water droplets on the inner wall of the tube cap.

[0041] (4) Add 200 μl of absolute ethanol, shake and mix well for 15 seconds, at this time flocculent precipitation may appear, and remove the water droplets on the inner wall of the tube cap...

Embodiment 2

[0058] Using MassARRAY molecular weight array technology (Li Handong et al., Application of matrix-assisted laser desorption / ionization time-of-flight mass spectrometry in microbial genome research. Journal of Jining Medical College. 2011, 63-67) to detect the genotypes of the local goat population in Enshi, Hubei Province , The genotype frequencies and gene frequencies of the goat population are shown in Table 1. The results showed that all genotypes of TLR2 gene were distributed in the population. Among the 200 individuals, there were 118 individuals with GG genotype, 62 individuals with GA genotype, and 20 individuals with AA genotype. The G allele frequency is 74.5%, and the A allele frequency is 25.5%. The G allele predominates.

[0059] Table 1 PCR--RFLP--BsaHI polymorphism of the present invention is distributed in Chinese native goats

[0060]

Embodiment 3

[0062] The Chinese native goat populations used in the association analysis between genotypes and anti-parasitic infection traits came from four natural villages in Enshi, Hubei Province. Rectal feces collection and ear tissue samples were collected from all individuals, and the preparation of genomic DNA samples from Chinese local goats was a routine method.

[0063] The traits analyzed were mainly anti-parasitic infection traits (ie presence or absence of parasite infection). According to the population structure of the collected samples, the applicant used the plink software (using the Case / Control model) to statistically analyze the genotype effect of the SNP site of the TLR2 gene and its relationship with the disease resistance index.

[0064] The association analysis between the polymorphic site at 557bp of TLR2 gene DNA fragment and the anti-parasitic infection traits of Chinese local goats was carried out. It was found that it was significantly related to the disease ...

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Abstract

The invention belongs to the technical field of preparation of molecular markers of livestock and specifically relates to preparation and an application of a molecular marker related to anti-infection of parasite of a goat. The molecular marker is obtained by cloning a goat TLR (Toll Like Receptor) 2 gene. The nucleotide sequence of the molecular marker is shown as a sequence table SEQ ID NO:1. A G / A mutant is located at the 557th basic group shown as the sequence table SEQ ID NO:1 and the mutant leads to RFLP (Restriction Fragment Length Polymorphism)-BsaHI polymorphism. The invention further discloses a preparation method of obtaining the molecular marker and an application of a polymorphism detection method, thereby providing a new molecular marker for correlation analysis of marker assisted selection of the goat.

Description

technical field [0001] The invention belongs to the technical field of animal molecular marker preparation, and in particular relates to the preparation and application of a molecular marker related to anti-parasitic infection traits of goats. Background technique [0002] Chinese goats have a long history of raising goats, and there are written records of raising goats as early as the Xia and Shang dynasties. More than a thousand years ago, goats were mainly raised in southern my country, and then gradually formed a scale. Over the past 30 years of reform and opening up, my country's goat industry has developed rapidly. However, due to the impact of species, diseases and ecological environment factors, its consumption is still low. At present, China's per capita mutton is less than 2.5kg. Parasitic disease is one of the important diseases that threaten the development of goat breeding industry, and it is also the most common disease in goats. After goats are infected with ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/11C12N15/10C12Q1/68
Inventor 赵书红付玉华曹建华吴震洋李新云李长春朱猛进余梅
Owner HUAZHONG AGRI UNIV