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Single nucleotide polymorphism (SNP) molecular marker related to porcine red blood cell quantitative trait after PolyI:C infection

A technology of molecular markers and red blood cells, which is applied in the direction of recombinant DNA technology, microbial detection/testing, DNA/RNA fragments, etc., can solve the problems of pig production performance decline, inability to completely control and eliminate infectious diseases, etc., and achieve high sensitivity and specificity good sex effect

Active Publication Date: 2018-09-07
HUAZHONG AGRI UNIV
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  • Description
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AI Technical Summary

Problems solved by technology

In pig production, diseases, especially viral infectious diseases, seriously threaten the health of pigs. Even though vaccination and drug treatment have played an important role, they cannot completely control and eliminate the occurrence and prevalence of infectious diseases (Sun Shouyong et al., Pig's Disease resistance and disease-resistant breeding[J]. Animal Science and Veterinary Medicine, 2005,22(12):60-61), in addition, the use of vaccines, while preventing diseases, will also make pigs in a highly activated state, so that the production performance of pigs will decline (Yang Jianhui et al., Analysis of the causes of diseases and epidemic prevention countermeasures in large-scale pig farms [J], Hunan Agricultural Science, 2011, 2011(1): 130-133)

Method used

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  • Single nucleotide polymorphism (SNP) molecular marker related to porcine red blood cell quantitative trait after PolyI:C infection
  • Single nucleotide polymorphism (SNP) molecular marker related to porcine red blood cell quantitative trait after PolyI:C infection
  • Single nucleotide polymorphism (SNP) molecular marker related to porcine red blood cell quantitative trait after PolyI:C infection

Examples

Experimental program
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Embodiment 1

[0025] Example 1: Genotyping Detection

[0026] (1) Using phenol extraction method to extract ear tissue DNA from Duroc×Erhualian F2 population

[0027] 1) Grind the ear tissue of Duroc×Erhualian F2 generation pig population (provided by Guangdong Huanong Wens Animal Husbandry Co., Ltd.) in liquid nitrogen, add an equal volume of 1×SET (1mL), proteinase K (10ng / mL) to a final concentration of 200ug / mL, then add 10% sodium dodecyl sulfate (SDS) to a final concentration of 0.5%, and shake well. Incubate the digestion overnight in a 55 °C water bath.

[0028] 2) Add an equal volume of Tris-saturated phenol to the digested tissue sample, slowly invert the centrifuge tube for 15 minutes, centrifuge in a low-temperature refrigerated centrifuge at 4°C and 11,000 rpm for 10 minutes, and carefully transfer the supernatant to another centrifuge tube. Mark the corresponding mark.

[0029] 3) Add an equal volume of phenol / chloroform / isoamyl alcohol (volume ratio 25:24:1), slowly inver...

Embodiment 2

[0039] Example 2: Application of CTTNBP2 molecular marker typing method in correlation analysis of pig immune traits

[0040] (1) Correlation analysis between CTTNB2 molecular marker typing results and immune traits

[0041] The experimental pigs used for the detection and analysis of the association between genotype and immune traits come from the F2 generation population (conventional breed) of the Duroc × Erhualian hybrid bred by Guangdong Huanong Wenshi Animal Husbandry Co., Ltd. The DNA used for genotyping is extracted from the ear sample of the F2 generation of Duroc×Erhualian cross (the “F2 generation of Duroc×Erhualian cross” referred to as “pig” in the text of the manual and the table) is used for routine blood testing Blood for flow cytometric analysis was collected from 35-day-old pigs 4 hours after polymyocyte (PolyI:C) inoculation. Using the method based on the single-marker association regression model, using the gender of the individual as a fixed effect, the G...

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Abstract

The invention belongs to the technical field of preparation of porcine molecular markers, and in particular relates to a single nucleotide polymorphism (SNP) molecular marker related to porcine red blood cell quantitative trait after PolyI:C infection. The molecular marker is cloned from a CTTNBP2 gene with an accession number DIAS0002844; the gene is subjected to somatotype screening by means ofa Porcine SNP60 BeadChip SNP technology, so that the SNP molecular marker related to the porcine red blood cell quantitative trait after PolyI:C infection is obtained; the nucleotide sequence of the marker is shown as SEQ ID NO. 1, and an A / G allelic mutation exists at the basic group of the 222nd position of the sequence; the mutation causes Porcine SNP60 BeadChip SNP typing polymorphism. The invention discloses a method for screening the molecular marker and application of the molecular marker in a correlation analysis method. The invention provides the novel SNP molecular marker for marker-assisted selection of porcine immune traits, particularly a red blood cell number.

Description

technical field [0001] The invention belongs to the technical field of preparation of porcine molecular markers, and in particular relates to SNP molecular markers related to porcine erythrocyte number traits after polymyocyte infection. The molecular marker described in the invention can be used for the prediction of the character of pig red blood cell number (RBC). Background technique [0002] In recent years, my country's animal husbandry has developed rapidly. According to data, my country's pig output value in 2016 was 1.2 trillion yuan, accounting for 43.2% of the animal husbandry output value (Huang Yueqin, Discussion on the current situation and development trend of my country's pig industry [J], Livestock Poultry Industry, 2017,28(8):102-102), my country's farming model has also changed from small-scale free-range farming to large-scale farming. In pig production, disease obviously becomes one of limiting factors for the development of breeding industry. According ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6837C12N15/11
CPCC12Q1/6837C12Q1/6888C12Q2600/124C12Q2600/156
Inventor 刘向东南九红赵书红李新云赵志超张杰朱猛进许月园胡明阳王昇
Owner HUAZHONG AGRI UNIV
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