Goose-origin gene RIG-I (retinoic acid-inducible gene-I) with anti-Newcastle disease virus activities and application thereof

An anti-Newcastle disease virus, gene technology, applied in antiviral agents, applications, gene therapy and other directions, can solve the problem of less research on antiviral immunity

Inactive Publication Date: 2013-06-26
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although geese, as waterfowl, have been reported to have Newcastle disease in recent years, compared with terrest

Method used

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  • Goose-origin gene RIG-I (retinoic acid-inducible gene-I) with anti-Newcastle disease virus activities and application thereof
  • Goose-origin gene RIG-I (retinoic acid-inducible gene-I) with anti-Newcastle disease virus activities and application thereof
  • Goose-origin gene RIG-I (retinoic acid-inducible gene-I) with anti-Newcastle disease virus activities and application thereof

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Experimental program
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Embodiment 1

[0034] 1. Determination of goose RIG-I gene sequence and construction of eukaryotic expression plasmid

[0035] 1.1 Primer design

[0036] According to the existing RIG-I sequences of humans, ducks and zebra finches on GenBank, overlap PCR amplification primers P1, P2, P3, and P4 were designed to amplify the RIG-I gene of goose by fusion PCR, wherein P1, P2 is the upstream and downstream primers of the first round of PCR, and P3 and P4 are the upstream and downstream primers of the second round of PCR. After the two rounds of PCR products are mixed, P1 and P4 are used to perform the third round of PCR to obtain the whole field sequence. According to the conserved region, the identification primers P5, P6, and CARD region amplification primers P7, P8 were designed.

[0037] Table 1 Primer sequences used in RIG-I gene related PCR amplification

[0038] Primer sequence P1L 5′-CGGCCGGCAGAGCCCAGCC-3′ P2R 5′-GTCTTAGCAAAGAGAAGAGGTGCGAGTCTGTGGGTTATAGC-3′ ...

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Abstract

The invention provides a goose-origin gene RIG-I (retinoic acid-inducible gene-I) with anti-Newcastle disease virus activities and an application thereof. According to the goose-origin gene RIG-I and the application of the goose-origin gene RIG-I, mRNA (Messenger Ribonucleic Acid) sequences of the goose-origin gene RIG-I are amplified based on a duck-origin gene RIG-I, and the homology of the goose-origin gene RIG-I and the duck-origin gene RIG-I is found to reach 93.8%; heterogenously-expressed gRIG-I is proved to be capable of expressing an anti-NDV (Newcastle disease virus) effect after over-expressing transfection cells of the full-length gRIG-I (gRIG-I full) and a gRIG-I CARD structural domain (gRIG-I CARD); the gRIG-ImRNA levels are detected by respectively infecting primary-culture geese embryo fibroblasts (GEF) and 2-week Yangzhou geese by utilizing three NDV viruses; the gRIG-ImRNA levels are found to averagely rise after the GEFs are infected by the NDV; and after being infected by the NDV, the gRIG-ImRNA levels in tissues of lungs and air sacs of the geese averagely rise, and the virus growths in in-vitro and in-vivo experiments are all restrained.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a goose-derived gene sequence with anti-Newcastle disease virus (Newcastle disease virus, NDV) activity, and the application of the gene. Background technique [0002] Newcastle disease is a highly contagious and fatal infectious disease to poultry. Since chickens are the most susceptible birds, the disease often causes huge losses to the poultry industry. The disease is caused by Newcastle disease virus (NDV), which is a member of the genus Mumps virus in the family Paramyxoviridae and is a single-stranded negative-strand non-segmented enveloped RNA virus. [0003] Pattern recognition receptors (pattern recognition receptors, PRR) are an important natural immune system of the host, which can recognize a variety of microbial components, such as viral nucleic acid, etc., quickly initiate a variety of signal cascade reactions, and synthesize a series of cytokines, including ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C07K14/465A61K48/00A61P31/14A01K67/027
Inventor 丁铲
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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