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Gene recombinant fowl influenza virus D3/F-R2/6 and its construction method

A technology of avian influenza virus and construction method, which is applied in the direction of virus/bacteriophage, DNA/RNA fragments, genetic engineering, etc., can solve difficult, costly, time-consuming and labor-intensive problems

Inactive Publication Date: 2006-02-15
YANGZHOU UNIV
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  • Abstract
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Problems solved by technology

[0004] Because there are many subtypes of influenza virus, and there are different mutant strains of the same subtype, it is very difficult to select the appropriate strain for the production of influenza vaccines.
In addition, some isolates cannot be multiplied to high enough titers, and laborious concentration is required to produce a vaccine with sufficient protection [Palese P, García-Sastre. Influenza A Vaccines: present and future. Clinical Investigation, 2002, 110: 9- 13.]; At the same time, due to the variability of influenza virus antigens, the matching of vaccines must be re-evaluated every year. This process is time-consuming and laborious

Method used

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  • Gene recombinant fowl influenza virus D3/F-R2/6 and its construction method
  • Gene recombinant fowl influenza virus D3/F-R2/6 and its construction method

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Embodiment Construction

[0101]The MPAIV and HPAIV strains selected in the present invention must have good immunity and high replication ability in chicken embryos. The present invention selects the MPAIV A / Chicken / Shanghai / F / 98 (H9N2) strain used as the H9N2 subtype vaccine, abbreviated as F strain, which is isolated and purified by the Key Open Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agriculture, Yangzhou University , was identified as H9N2 subtype by the National Influenza Center, and the full gene sequence was registered in GenBank with the accession number AY253750-AY253756 [Lu Jianhong, Liu Xiufan, Shao Weixing, et al. Determination of the full-length sequence of the H9N2 subtype AIV genome and genetic analysis of each gene. Acta Microbiology, 2003, 43(4): 434-41.]. Select the HPAIV A / Duck / Huadong / D3 / 00 (H5N1) strain, referred to as the D3 strain, which was isolated and purified by the Key Open Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agr...

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Abstract

The invention provides an avian influenza virus D3 / F-R2 / 6*reassortment and construction method thereof, which is related to the field of reversal genetic technology and comprises, reassorting the six internal genes of the MPAIV chicken embryonic highly adaptive strain A / Chicken / Shanghai / F / 98(H9N2) PB2, PB1, PA, NP, M, NS and the NA gene and attenuate HA geine of the HPAIV epidemigenic strain A / Duck / Huadong / D3 / 00 (H5N1), thus constructing an avian influenza virus D3 / F-R2 / 6*reassortment, which can be used for manufacturing vaccine for treating avian influenza.

Description

technical field [0001] The present invention relates to the application of reverse genetic technology. In particular, two external genes of a strain of H5N1 subtype avian influenza virus (abbreviation: H5N1 subtype AIV) and 6 internal genes of a strain of H9N2 subtype avian influenza virus (abbreviation: H9N2 subtype AIV) were constructed into 8 All genes are derived from genetically reassorted strains of AIV for use in the manufacture of vaccines. Background technique [0002] The reverse genetic manipulation technology of RNA viruses refers to the construction of infectious molecular clones of RNA viruses in vitro, that is, the reverse transcription of viral genome RNA into cDNA, and various in vitro artificial manipulations on the DNA molecular level. A technique in which various accessory proteins are used to assemble new RNA viruses. Since the "rescued" RNA virus is derived from cDNA clones, various in vitro artificial manipulations can be performed on the RNA virus g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/34C12N15/62A61K39/12
Inventor 刘秀梵石火英卢建红张小荣
Owner YANGZHOU UNIV
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