Molecular marker and detection method of thermal stress endurance of holstein

A technology of Holstein cattle, heat stress, applied in the biological field, can solve problems such as deficiency

Active Publication Date: 2019-08-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are few studies on DNA molecular markers of heat stress tolerance of dairy cows at home and abroad, and there is still a lack of quantitative trait loci associated with production

Method used

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  • Molecular marker and detection method of thermal stress endurance of holstein
  • Molecular marker and detection method of thermal stress endurance of holstein
  • Molecular marker and detection method of thermal stress endurance of holstein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Example 1. Genome-wide association analysis of rectal temperature of Holstein cattle under heat stress conditions

[0107] 1. Determination of rectal temperature

[0108] From 2013 to 2015, the rectal temperature of Holstein lactating cows under heat stress in 9 pastures in Beijing was measured every summer (June to August), a total of 6,446 cows. Measuring time: 7:00 to 10:00 in the morning and 14:00 to 17:00 in the afternoon (according to NY / T 2363-2013, when the average temperature and humidity index of the barn is 72≤THI≤79, Holstein cattle In a state of mild heat stress; when the average temperature and humidity index of the barn is 7888, the Holstein cattle are in a severe heat stress state. Stress state), each cow measured rectal temperature twice a day, recorded as rectal temperature in the morning and rectal temperature in the afternoon, and measured for two consecutive days. Measuring method: When measuring, the cow is under the neck yoke, and the electronic...

Embodiment 2

[0120] Example 2 Screening and polymorphism mining of candidate genes related to heat stress tolerance

[0121] In order to find candidate genes that may be related to heat stress tolerance, the 200kb regions near the significant SNPs were selected as candidate regions, and the gene screening was carried out. A total of 5 candidate genes that may be related to EBV in the rectal temperature in the morning were found (Table 1). After reviewing the literature, it was found that the protein encoded by the PHRF1 gene is rich in the Ser / Arg domain, which binds to the C-terminus of the large subunit of RNA polymerase II, and can promote genomic DNA by regulating non-homologous end joining after DNA damage. For completeness, we therefore speculate that PHRF1 is the most likely functional candidate gene. In order to further verify the possibility of the PHRF1 gene as a candidate gene, its polymorphism was screened and its association with traits was analyzed.

[0122] 1. Genomic DNA ...

Embodiment 3

[0152] Example 3. SNPs group typing and its correlation analysis with various physiological indicators and milk production performance EBVs of Holstein cattle under heat stress

[0153] Randomly select 1,665 Holstein lactating cows with clear pedigree, complete phenotype and DHI records from Beijing area as the experimental group, collect 10 mL of anticoagulated blood intravenously, and then take 800 μL of blood, and use the blood genome extraction kit (article number: DP318-03, Tiangen Biochemical Technology (Beijing) Co., Ltd.), DNA extraction was carried out according to the instructions. Use NanoDrop2000 ultraviolet spectrophotometer and 1% agarose gel electrophoresis (130V, 20min) to detect the quality of DNA extraction, and store the qualified DNA in a -20°C refrigerator for future use.

[0154] 1. Establishment of a large group typing method for SNPs of the PHRF1 gene

[0155] Each individual SNP genotype was obtained using competitive allele-specific PCR (KASP). Comp...

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Abstract

The invention discloses a molecular marker and a detection method of thermal stress endurance of a holstein. The invention provides one of the following application adopted to detect whether deoxyribonucleotide on a 2029th site of a cDNA sequence of a PHRF1 gene of a holstein genome is T, C or a substance of T and C: (A) verifying and assisting in verifying thermal stress endurance of the holstein; (B) breeding a holstein herd with relatively high thermal stress endurance. The molecular marker and detection method disclosed by the invention are simple in operation, automatic machine detectioncan be conducted to obtain the type of an individual gene when only an individual genome is obtained, so that thermal resistance of the individual is known. The cost is low, the accuracy is high, andboth time and energy are save. In total, when a CC gene type on a T2029C site of the PHRF1 gene is adopted to select the thermal endurance of the holstein, the thermal endurance of the holstein is improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a molecular marker and a detection method of Holstein cattle heat stress tolerance. Background technique [0002] At present, the continuous warming of the global climate poses a threat to human development, animal survival and ecological balance. The increasingly serious heat stress affects the production performance, reproductive performance and disease resistance of livestock and poultry, and restricts the sustainable development of animal husbandry. The sweat glands of dairy cows are underdeveloped, their heat dissipation ability is poor, and a lot of heat is also generated during milk production, which makes dairy cows particularly sensitive to heat stress. According to reports, the decline in cow milk production caused by heat stress can cause the entire US dairy industry to lose $1.2 billion per year, and the German dairy industry also loses more than 10% of the annual econom...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/124C12Q2600/158
Inventor 王雅春刘嘉莉胡丽蓉罗汉鹏初芹徐青
Owner CHINA AGRI UNIV
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