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CD163 gene fragment clone related to pig immune trait and use thereof as molecular marker

A technology of molecular markers and gene fragments, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of no report on porcine CD163 gene polymorphism, no research on porcine CD163 gene molecular markers, etc.

Inactive Publication Date: 2009-08-12
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, at present, the research on CD163 gene at home and abroad is mainly focused on the function of anti-inflammatory response, and the polymorphism of porcine CD163 gene has not been reported. So far, no molecular markers related to porcine CD163 gene and porcine immunity have been studied.

Method used

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  • CD163 gene fragment clone related to pig immune trait and use thereof as molecular marker
  • CD163 gene fragment clone related to pig immune trait and use thereof as molecular marker
  • CD163 gene fragment clone related to pig immune trait and use thereof as molecular marker

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Experimental program
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Effect test

Embodiment 1

[0031] 1. Cloning of partial fragments of CD163 gene and screening of mutation sites

[0032] 1. Primer Design

[0033] Using the primer design software Primer 5.0, the amplification primers containing part of the coding region were designed using the porcine CD163 gene mRNA sequence (Genebank accession number NM_213976.1) as a template. The primers were synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd. The DNA sequence of the primer pair is as follows:

[0034] Forward primer: 5′-TGGTCAACTTCGCCTGGT-3′,

[0035] Reverse primer: 5'-AGCACGTCACAGCAGCAT-3'.

[0036] 2. Purification and sequencing of PCR products

[0037] (1) PCR amplification

[0038]PCR amplification reaction system 20ul, including 50ng of cDNA, 1×PCR buffer, 2.5mM MgCl, 0.2mM dNTP, 10pmol ofeach primer and 1 U Ampli Taq DNA polymerase (Shanghai Sangon Bioengineering Technology Co., Ltd.). The PCR reaction conditions were as follows: the PCR reaction program was: 95°C for 5 minutes; 35 cycle...

Embodiment 2

[0079] Application of the CD163 molecular marker typing method established in the present invention in the correlation analysis of immune traits

[0080] (1) Immune traits associated with CD163 gene 2537 G-A (HpaII-RFLP) molecular marker

[0081] (1) The molecular marker 2537 G-A genotype was detected by CRS-PCR-HpaII-RFLP and the correlation analysis was carried out. The results showed that this molecular marker conformed to the Hardy-Weinberg equilibrium law in the detection group, and it had a positive effect on the reproductive and respiratory syndrome of 0-day-old pigs. Virus (PRRS) antibody detection value, platelet distribution width, platelet volume, ratio of large platelets, platelets and white blood cells were significantly affected. The specific results are shown in Table 1:

[0082] Table 1 The least squares mean of immune traits associated with different genotypes of CD163 gene 2537G-2537A

[0083]

[0084] P<0.05 means significant difference; P<0.01 means ex...

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Abstract

The invention belongs to the technical field of preparing livestock molecular markers, and in particular relates to preparation and application of a molecular marker applied as porcine marker for assisted selection and relevant to porcine immunity. The molecular marker is obtained by cloning a porcine CD163 gene, and a mRNA sequence of the molecular marker is shown in a sequence table SEQ ID NO:1. A 2439bp locus of the sequence table SEQ ID NO:1 has an A-to-G base mutation; a 2454bp locus of the sequence has a G-to-C base mutation; a 2537bp locus of the sequence has a G-to-A base mutation; and a 2685bp locus of the sequence has an A-to-C base mutation. The invention also discloses a primer for amplifying partial coding regions of the CD163 gene and a method for detecting a 2537bp G-to-A molecular marker and a 2685bp A-to-C molecular marker. The invention provides two novel molecular markers for porcine marker-assisted selection.

Description

technical field [0001] The invention belongs to the technical field of livestock molecular marker (genetic marker) preparation, and in particular relates to the cloning and application of a molecular marker related to pig immune traits used as pig marker-assisted selection. Background technique [0002] Porcine Reproductive and Respiratory Syndrome PRRS (Porcine Reproductive and Respiratory Syndrome, also known as "pig blue ear disease" due to cyanosis of pig ears and so on) is caused by porcine reproductive and respiratory syndrome virus (PRRSV) infection, is One of the most important diseases afflicting pig production, causing serious economic losses worldwide. In recent years, due to severe outbreaks of PRRS, avian influenza and foot-and-mouth disease worldwide, drug and harmful substance residues affect animal food safety and the environment, animal disease resistance genetic breeding has received more and more attention, and has become an important goal of animal breedi...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12Q1/68C12P19/34
Inventor 张淑君陈焕春童勤赵书红杨利国赵俊龙方六荣王振骅张立斌刘向东戴益刚
Owner HUAZHONG AGRI UNIV
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