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Unmethylated CpG contained DNA sequence (CpG-DNA) and its application

A DNA sequence, unmethylated technology, applied in the field of immunology, can solve the problems of no related reports, achieve the effect of inhibiting virus infection and enhancing specific immune effect

Inactive Publication Date: 2014-02-05
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the CpG-DNA that can induce significant antiviral infection in flounder

Method used

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  • Unmethylated CpG contained DNA sequence (CpG-DNA) and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The flounder specific CpG-DNA sequence has the base sequence in SEQ ID No.1 of the sequence list.

[0019] The sequence listing SEQ ID No.1 is:

[0020] GGCGCGCGTCGCGCGCTA

[0021] (a) Sequential features:

[0022] ●Length: 18

[0023] ●Type: Nucleotide sequence

[0024] ●Chain type: single chain

[0025] ●Topological structure: linear

[0026] (b) Molecule type: DNA

[0027] (c) Assumption: No

[0028] (d) Antisense: No

[0029] (e) Original Source: Artificial Design

[0030] Example 1

[0031] Application of CpG-DNA as antiviral factor

[0032] 1) Preparation of virus suspension. Swelling cell virus RBIV-C1 (see Zhang M, Xiao Z, Hu Y, Sun L.Characterization of a megalocytivirus from cultured rock bream, Oplegnathus fasciatus (Temminck&Schlege), in China.Aquac Res.2012; 43: 556–64) were diluted to 5x10 in PBS 6 copies / ml is the virus suspension.

[0033] The composition of PBS is by weight percentage: 0.8%NaCl, 0.02%KCl, 0.358%Na 2 HPO 4 .12H 2 O, 0.02...

Embodiment 2

[0036] Application of CpG-DNA as Vaccine Adjuvant

[0037] Step 1) Vaccine Immunization Injection

[0038] The DNA vaccine plasmid pCN86 (see Zhang M, Hu Y, Xiao Z, Sun Y, Sun L. Construction and analysis of experimental DNA vaccines against megalocytivirus. Fish Shellfish Immunol. 2012; 33:1192–8. for the construction process) in PBS Diluted to 200ug / ml. Dilute CpG-DNA to 200ug / ml in PBS. Mix equal volumes of pCN86 and CpG-DNA diluent, that is, pCN86+CpG-DNA mixed solution. Mix equal volumes of pCN86 and PBS to obtain pCN86 vaccine solution. 15 flounder (same as above) were randomly divided into 3 groups, 5 in each group. These 3 groups are named A, B and C respectively. Inject 0.1ml of pCN86 vaccine liquid into each fish in group A, inject 0.1ml pCN86+CpG-DNA mixture into each fish in group B, and inject 0.1ml PBS into each fish in group C (control group).

[0039] Step 2) For the preparation of virus suspension, refer to the corresponding records in Example 1.

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Abstract

The invention relates to the field of immunology, specifically to an unmethylated CpG contained DNA sequence which has immune stimulation effect in Paralichthys olivaceus and its application in anti-virus. The unmethylated CpG contained DNA sequence (CpG-DNA) is a CpG-DNA sequence which has immune stimulation effect in Paralichthys olivaceus. The CpG-DNA sequence is as shown in the base sequence of sequence table SEQ ID No.1. The CpG-DNA provided by the invention can defense the infection of megalocytivirus, can be used as a vaccine adjuvant and can enhance specific immune effect of vaccine.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a DNA sequence containing unmethylated CpG with immune stimulating effect in flounder and its application in anti-virus. Background technique [0002] CpG-DNA is a DNA sequence containing unmethylated CpG, which is a characteristic of prokaryotic DNA sequences. In the course of evolution, vertebrates have acquired an immune recognition system against CpG-DNA to make a protective immune response to microbial pathogen invasion, including activating Th1-induced cellular immunity and Humoral immunity, etc. Studies have shown that CpG-DNA has species specificity, the effective CpG-DNA sequences are different in different species, and the immune response induced by different CpG-DNA in the same species is also different. Flounder is an important marine fish cultured in my country, and its aquaculture industry has been continuously affected by microbial diseases in recent years. Although th...

Claims

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Application Information

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IPC IPC(8): C12N15/117A61K31/711A61K39/39A61P31/20
Inventor 孙黎周志侠孙铂光
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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