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Fusion gene for expressing Fc fragment of chicken IgY molecule and avian influenza virus antigen and application of same

An avian influenza virus and gene fusion technology, applied in the field of genetic engineering, can solve the problems of low overall efficiency, low proportion of M cells, increase the efficiency of mucosal vaccines, etc., achieve low cost, efficient transportation, and realize the effect of preventing avian influenza

Active Publication Date: 2015-08-12
CHINA AGRI UNIV
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Problems solved by technology

However, the proportion of M cells in mucosal epithelial cells is extremely low (only 1 M cell in 10 million epithelial cells), which makes the overall efficiency of antigen uptake by M cells low. New strategies must be adopted to increase the efficiency of mucosal vaccines

Method used

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  • Fusion gene for expressing Fc fragment of chicken IgY molecule and avian influenza virus antigen and application of same
  • Fusion gene for expressing Fc fragment of chicken IgY molecule and avian influenza virus antigen and application of same
  • Fusion gene for expressing Fc fragment of chicken IgY molecule and avian influenza virus antigen and application of same

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Embodiment 1

[0028] In this embodiment, Eimeria mitis is used as a carrier to fuse and express the Fc fragment of chicken IgY molecule and the envelope protein HA1 of avian influenza virus H9N2. The immune response to influenza-specific antigens can reduce the titer of avian influenza virus in infected chickens by 81%.

[0029] 1. Carrier construction

[0030] The total RNA of chicken and avian influenza virus H9N2 (isolated from sick chickens with symptoms of avian influenza in a poultry farm) were extracted by the traditional Trizol method, and corresponding primers were designed to amplify the HA1 gene fragment and the H9N2 subtype of avian influenza virus. Fc fragment of chicken IgY molecule. Further, the HA1 gene and the Fc gene fragments were connected by a bridge PCR method to form a fusion gene HA1-Fc, the nucleotide sequence of which is shown in SEQ ID No.1. Afterwards, subclone HA1-Fc onto the cloning vector, digest the correct HA1-Fc gene identified by sequencing with restrict...

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Abstract

The invention relates to a fusion gene for expressing Fc fragment of chicken IgY molecules and avian influenza virus antigen and an application of the same. The fusion gene is constructed by means of an HA1 gene fragment of avian influenza virus H9N2 sub-type and the Fc gene fragment of the chicken IgY molecule and then is transfected to eimeria mitis to obtain transgenic coccidium. The transgenic coccidium is inoculated through an oral method for enhance immune reaction aiming to an avian influenza specific antigen in chickens. The application is simple to carry out, is low in cost and has an excellent application prospect.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a fusion gene expressing chicken IgY molecule Fc fragment and avian influenza virus antigen and its application. Background technique [0002] The mucosa is the main site for most pathogens to invade the host, so targeting foreign antigens to the mucosa to stimulate a specific mucosal immune response is an important strategy to resist pathogen invasion. But the tight junctions between epithelial cells on the mucosal surface make penetration of molecules difficult. One solution is to target mucosal antigens to M cells in the intestinal epithelium -- a specialized type of epithelial cell that takes up antigens at the interface between the mucosal surface and the external environment. . Once the antigen is taken up by M cells, it can rapidly migrate to the base of epithelial cells and interact with Peyer's lymph nodes. However, the proportion of M cells in mucosal epithelial cel...

Claims

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Application Information

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IPC IPC(8): C12N15/62C12N15/85C12N15/66C12N1/11A61K48/00A61K35/68A61P31/16
Inventor 索勋刘贤勇秦梅汤新明索静霞
Owner CHINA AGRI UNIV
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