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H9N2 subtype AIV (H9N2 Subtype Avian Influenza Viruses) inhibitor and application thereof

A technology of avian influenza virus and inhibitors, applied in antiviral agents, genetic material components, recombinant DNA technology, etc., can solve the problems of relatively few researches on miRNA functions

Active Publication Date: 2019-12-20
POULTRY INST SHANDONG ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, more than 800 miRNAs have been found in chickens, but there are relatively few functional studies of these miRNAs

Method used

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  • H9N2 subtype AIV (H9N2 Subtype Avian Influenza Viruses) inhibitor and application thereof
  • H9N2 subtype AIV (H9N2 Subtype Avian Influenza Viruses) inhibitor and application thereof
  • H9N2 subtype AIV (H9N2 Subtype Avian Influenza Viruses) inhibitor and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Expression detection of gga-miR-30a-3p in DF-1 cells infected with H9N2 subtype avian influenza virus

[0021] (1) After infecting DF-1 cells with H9N2 subtype avian influenza virus at MOI=0.5, collect DF-1 cells at different time points (0 h, 6 h, 12 h, 24 h and 36 h) and extract total RNA , the expression of gga-miR-30a-3p was detected by fluorescent quantitative PCR, and the results showed that the expression level of gga-miR-30a-3p decreased continuously with the prolongation of infection time, and the expression level reached the lowest level at 24h after infection Level( figure 1 ).

[0022] (2) After infecting DF-1 cells with H9N2 subtype avian influenza virus at different doses (MOI=0.1, 0.5, 1, 1.5), the DF-1 cells were collected, total RNA was extracted, and gga-miR- The expression of 30a-3p, the results showed that with the increase of virus infection dose, the expression level of gga-miR-30a-3p was continuously down-regulated ( figure 2 )

Embodiment 2

[0024] Using gga-miR-30a-3p to inhibit the proliferation of H9N2 subtype avian influenza virus in DF-1 cells

[0025] (1) Synthesize the mimics (mimics) of gga-miR-30a-3p and the negative control NC, and different concentrations (10 nm, 20 nm, 40 nm, 80 nm) of the mimics / NC of gga-miR-30a-3p were respectively DF-1 cells were transfected, and 24 h after transfection, the cells were collected, and the total RNA was extracted by Trizol reagent (Invitrogen Company), and cDNA was synthesized by reverse transcription using the Thermo reverse transcription kit, and gga-miR-30a was detected by fluorescent quantitative PCR - the expression level of 3p;

[0026] (2) Before inoculating H9N2 subtype avian influenza virus (isolated by conventional methods), place the virus stock solution on ice and slowly thaw it. After 24 hours of transfection of DF-1 cells, use MOI=0.5 H9N2 subtype Cells were infected with avian influenza virus. After 24 hours of infection, the cells were collected, and...

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Abstract

The invention relates to the technical field of animal immunization and the technical field of biological medicine, in particular to chicken gga-miR-30a-3p as an H9N2 subtype AIV (H9N2 Subtype Avian Influenza Viruses) inhibitor and application thereof. The inhibitor is gga-miR-30a-3p mimics, and the sequence is shown as SEQ ID NO.1. The invention provides a design target for novel prevention and control medicine research and development of H9N2 subtype AIV. The invention also provides application of artificially synthesized gga-miR-30a-3p as medicine or a reagent for preventing and treating the H9N2 subtype AIV.

Description

technical field [0001] The invention relates to the technical field of animal immunity and the technical field of biomedicine, in particular to chicken gga-miR-30a-3p as an H9N2 subtype avian influenza virus inhibitor and its application. Background technique [0002] Avian influenza is a disease syndrome caused by Avian influenza virus (AIV) infecting poultry, including diseases such as respiratory tract, digestive tract, egg production decline or immunosuppression, which can be divided into highly pathogenic avian influenza , low pathogenic avian influenza and apathogenic avian influenza. Since my country first isolated H9N2 subtype avian influenza viruses (H9N2 subtype AIV) in Guangdong Province in 1994, H9N2 subtype avian influenza virus is still the main low-pathogenicity bird flu virus in chicken flocks in my country. Influenza subtype. The H9N2 subtype avian influenza virus has brought great harm to the poultry industry. The virus not only causes poultry respiratory ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/87A61K48/00A61K31/7105A61P31/16
CPCC12N15/1131C12N15/87A61K31/7105A61P31/16C12N2310/141
Inventor 孟凯于江王友令袁小远张玉霞杨金兴
Owner POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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