MTHFR (methylenetetrahydrofolate reductase) gene fragment as SNP (single-nucleotide polymorphism) molecular marker associated with sow foot-leg bone density

A technology of molecular markers and bone density, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of high price, limited application, rare research on livestock bones, etc., and achieve high sensitivity and good specificity

Active Publication Date: 2018-08-17
HUAZHONG AGRI UNIV
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  • Claims
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AI Technical Summary

Problems solved by technology

It can be seen that bone density is an important indicator in the study of animal bone strength, but due to the limitations of various production conditions, people's research on livestock bones is very rare
[0004] Bone mineral density, that is, the content of bone mineral per unit volume, the unit is g/cm3, common bone density measurement methods include radioabsorption method, dual-energy X-ray measurement method, quantitative computed tomography technology and axial ultrasonic transmission technology, etc. (Cao Jian, Application evaluation of bone mineral density (BMD) measurement[J], Frontiers of Medicine, 2013(2)), among which dual-energy X-ray measur

Method used

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  • MTHFR (methylenetetrahydrofolate reductase) gene fragment as SNP (single-nucleotide polymorphism) molecular marker associated with sow foot-leg bone density
  • MTHFR (methylenetetrahydrofolate reductase) gene fragment as SNP (single-nucleotide polymorphism) molecular marker associated with sow foot-leg bone density
  • MTHFR (methylenetetrahydrofolate reductase) gene fragment as SNP (single-nucleotide polymorphism) molecular marker associated with sow foot-leg bone density

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Experimental program
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Effect test

Embodiment 1

[0024] Example 1: Genotyping Detection

[0025] (1) Using the kit method to extract ear tissue DNA from bone density-related populations

[0026] 1) Cut the ear-like tissues of bone density-related populations (breeds: Large White pig and Landrace pig, provided by Guangxi Yangxiang Co., Ltd.) with ophthalmic scissors until it becomes a paste, add 200ulGA (included in the kit), and shake Mix well; then put it into a 56°C water bath for overnight digestion.

[0027] 2) Add 200ul buffer GB (included with the kit) to the digested tissue sample, mix thoroughly by inversion, and place at 70°C for 10 minutes, the solution should become clear, and centrifuge briefly to remove water droplets on the tube wall.

[0028] 3) Add 200ul of absolute ethanol, shake and mix well for 15 seconds, at this time flocculent precipitation may appear, briefly centrifuge to remove water droplets on the tube wall.

[0029] 4) Add the solution and flocculent precipitate obtained in the previous step int...

Embodiment 2

[0036] Example 2 Application of the Molecular Marker Typing Method of the Present Invention in Correlation Analysis of Pig Limb and Hoof Bone Density Traits (1) Correlation Analysis between Molecular Marker Typing Results and Limb and Hoof BMD Traits

[0037] The experimental pigs used for the correlation detection and analysis of genotype and bone density traits of limbs and hooves come from purebred sow groups bred by Guangxi Yangxiang Co., Ltd., including Large White pigs and Landrace pigs (conventional breeds). The DNA used for genotyping is extracted from the ear samples of purebred Large White and Landrace ("Purebred Large White and Landrace" referred to as "pig" in the text of the manual and tables for short). Using the method based on the single-marker association regression model, using the gender of the individual as a fixed effect, the GWAS analysis was performed using the GenABEL software package in the R statistical environment.

[0038] The specific regression an...

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Abstract

The invention belongs to the technical field of animal molecular marking and particularly relates to MTHFR (methylenetetrahydrofolate reductase) gene fragment as an SNP (single-nucleotide polymorphism) molecular marker associated with sow foot-leg bone mineral density. The molecular marker is cloned from the MTHFR gene fragment having the register number of M1GA0008657; the gene fragment is subjected to typing screening via a gene chip technology so as to obtain the molecular marker associated with sow foot-leg bone mineral density; a nucleotide sequence of the molecular marker is shown as inSEQ ID NO: 1; base 51 of the nucleotide sequence has an A/C allele mutation; when nucleotide 51 in the SEQ ID NO: 1 is A, sow leg-foot bone has higher bone mineral density. Disclosed a method of screening molecular markers associated with sow pig-leg bone mineral density, as well as application of association analytical method. The novel SNP molecular marker is provided for marking-assisted selection of pig leg-foot compactness traits, particularly bone mineral density.

Description

technical field [0001] The invention relates to the technical field of preparation of pig molecular markers, in particular to the use of MTHFR gene fragments as SNP molecular markers related to bone density of sows' limbs and hooves. The molecular marker of the invention is cloned from the MTHFR gene, and can be used to predict the bone density traits of pig limbs and hooves. Background technique [0002] With the development of science and technology, the pig breeding industry has made significant breakthroughs in genetic breeding, nutritional regulation, and disease prevention and control. The lean meat rate, feed-to-meat ratio, and growth rate of commercial pigs have all been greatly improved. However, with intensive farming comes the increasingly prominent problem of limbs and feet. For intensive production, the range of activities of pigs is limited. In addition, the design of pens (slatted floors) has exacerbated the occurrence of pigs' limbs and feet. It has been rep...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 刘向东冯帅飞赵书红李新云刘小磊尹立林唐振双叶灯灯马官军
Owner HUAZHONG AGRI UNIV
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