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WNT2 (wingless-type MMTV integration site family, member 2) gene fragment as molecular marker influential on porcine red blood cell number trait

A technology of molecular markers and gene fragments, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc., to achieve good specificity and high sensitivity

Inactive Publication Date: 2018-08-17
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, food safety problems caused by drug residues have been plagued by agricultural and animal husbandry enterprises. Disease-resistant breeding can solve this problem and allow the cost of disease-resistant management to be shared equally

Method used

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  • WNT2 (wingless-type MMTV integration site family, member 2) gene fragment as molecular marker influential on porcine red blood cell number trait
  • WNT2 (wingless-type MMTV integration site family, member 2) gene fragment as molecular marker influential on porcine red blood cell number trait
  • WNT2 (wingless-type MMTV integration site family, member 2) gene fragment as molecular marker influential on porcine red blood cell number trait

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1: Genotyping Detection

[0025] (1) Using phenol extraction method to extract ear tissue DNA from Duroc×Erhualian F2 population

[0026] 1) Grind the ear tissue samples of the Duroc × Erhualian F2 generation pig population (group samples provided by Guangdong Huanong Wens Animal Husbandry Co., Ltd.) in liquid nitrogen, add an equal volume of 1 × SET (1 mL), 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.

[0027] 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.

[0028] 3) Add an equal volume of phenol / chloroform / isoamyl alcohol (volu...

Embodiment 2

[0038] Example 2: Application of the WNT2 molecular marker typing method in the correlation analysis of pig immune traits

[0039] (1) Correlation analysis between WNT2 molecular marker typing results and immune traits

[0040] 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 Duroc×Erhualian crossbred by Guangdong Huanong Wen's Animal Husbandry Co., Ltd. The DNA used for genotyping is extracted from the ear tissue of the F2 generation of Duroc × Erhualian cross ("Duroc × Erhualian hybrid F2 generation" referred to as "pig" in the text of the manual and the table) is used for blood Blood samples for routine testing and flow cytometry were collected from 35-day-old piglets 4 hours after inoculation with polymyocytes (PolyI:C). Using the method based on the single-marker association regression model, using the gender of the individual as a fixed effect, t...

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Abstract

The invention belongs to the technical field of animal molecular marking and screening and particularly relates to a WNT2 (wingless-type MMTV integration site family, member 2) gene fragment as a molecular marker influential on porcine red blood cell number trait. The molecular marker is cloned from WNT2 gene and is a fragment of WNT2 gene; the gene fragment is subjected to typing screening via agene chip technology to obtain the molecular marker associated with porcine red blood cell number; a nucleotide sequence of the molecular marker is shown as in SEQ ID NO: 1; base 24 of the nucleotidesequence has a T / C allele mutation; when the nucleotide 24 in the SEQ ID NO: 1 is C, it is determined that the number of red blood cells is higher. The novel SNP molecular marker is provided for marking assisted selection of porcine immunity traits, particularly, red blood cell number.

Description

technical field [0001] The invention belongs to the technical field of animal molecular marker screening, and specifically relates to a WNT2 gene fragment as a molecular marker affecting the number traits of pig red blood cells. The molecular marker of the invention is cloned from the WNT2 gene. The molecular marker can be used for predicting the character of pig red blood cell number (RBC). Background technique [0002] my country has a large population and likes to eat pork, and the consumption of pork is large, accounting for half of the world's total (Liu Wentao et al., 2016). According to the data from the National Bureau of Statistics, in 2017, my country's pork output was 53.4 million tons, imported pork was 1.217 million tons, and the import value was as high as 15.06 billion yuan. Gradually transform into high-efficiency and low-cost large-scale farming, and the free-range farming model is gradually eliminated (Deng Qifeng et al., 2017). Large-scale farming pays at...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 刘向东王梦芬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