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A goose-derived gene rig-i with anti-Newcastle disease virus activity and its application

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

Inactive Publication Date: 2016-02-03
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although geese, as waterfowl, have been reported to have Newcastle disease in recent years, compared with terrestrial poultry, they have stronger resistance to NDV, but there are few studies on their antiviral innate immunity.

Method used

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  • A goose-derived gene rig-i with anti-Newcastle disease virus activity and its application
  • A goose-derived gene rig-i with anti-Newcastle disease virus activity and its application
  • A goose-derived gene rig-i with anti-Newcastle disease virus activity and its application

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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, overlapPCR amplification primers P1, P2, P3, and P4 were designed to amplify the RIG-I gene of goose by fusion PCR, wherein P1, P2 It 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 for 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-derived gene RIG-I with anti-Newcastle disease virus activity, which is based on the duck-derived RIG-I gene, amplifies the goose-derived RIG-I gene mRNA sequence, and finds that the homology between the two reaches 93.8%. The present invention combines full-length gRIG-I (gRIG-I?full) and gRIG-I? The CARD domain (gRIG-I? CARD) was overexpressed in transfected cells, confirming that exogenously expressed gRIG-I can exert anti-NDV effect. Three kinds of NDV viruses were used to infect primary goose embryo fibroblasts (GEF) and 2-week-old Yangzhou geese to detect gRIG-I? mRNA levels; found gRIG-I after GEF infection with NDV? mRNA levels were up-regulated. gRIG-I in lung and air sac tissues of geese infected with NDV? mRNA levels were upregulated, and virus growth was inhibited in both in vitro and in vivo experiments.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a goose-derived gene sequence with anti-Newcastle disease virus (Newcastlediseasevirus, 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 paramyxoviridae mumpsvirus genus and is a single-stranded negative-strand non-segmented enveloped RNA virus. [0003] Pattern recognition receptors (pattern recognition receptors, PRR) is 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 type I interferen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C07K14/465A61K48/00A61P31/14A01K67/027
Inventor 丁铲孙英杰仇旭升谭磊宋翠萍于圣青
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI