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Newcastle disease virus rNDV-H52AH9 and construction method and application thereof

A newcastle disease virus, rndv-h52ah9 technology, applied in the direction of virus/bacteriophage, recombinant DNA technology, microorganism-based methods, etc., can solve the problems of complex virus epidemic situation and many vaccines

Inactive Publication Date: 2013-05-01
YANGZHOU UNIV
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Problems solved by technology

[0012] Therefore, the first object of the present invention is to provide a kind of recombinant vaccine strain that takes Newcastle disease virus as carrier to co-express H5 and H9 subtype avian influenza virus hemagglutinin protein, thereby solves the current gene VII type Newcastle disease virus, H5 and H9 The prevalence of H9 subtype avian influenza virus is complicated, and the vaccines used are many and complicated, which has the effect of preventing multiple diseases with one shot

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  • Newcastle disease virus rNDV-H52AH9 and construction method and application thereof
  • Newcastle disease virus rNDV-H52AH9 and construction method and application thereof
  • Newcastle disease virus rNDV-H52AH9 and construction method and application thereof

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Embodiment Construction

[0032] The recombinant vaccine strain rNDV-H co-expressing H5 and H9 subtype avian influenza virus hemagglutinin proteins constructed by the present invention using Newcastle disease virus as a carrier 5 2AH 9 On October 11, 2012, it was deposited in the General Microbiology Center of China Microbiological Culture Collection Management Committee. The address of the preservation unit is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences; its preservation number CGMCC No: 6654 , the classification is named: Newcastle Disease Virus (Newcastle Disease Virus), and its length is: 18648bp.

[0033] Construction step 1: Construction of full-length expression clones co-expressing H5 and H9 subtype avian influenza virus hemagglutinin proteins using attenuated Newcastle disease virus AI4 as a vector

[0034] Biomaterial preparation:

[0035] Attenuated full-length expression clone pNDV / AI4 of Newcastle disease virus JS / ...

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Abstract

The invention relates to a Newcastle disease virus rNDV-H52AH9, accession number CGMCC (China General Microbiological Culture Collection Center) No: 6654. An attenuated strain AI4 of the Newcastle disease virus is used as a carrier, HA genes of H5 and H9 subtype avian influenza virus are connected in series through FMDV (Foot and mouth disease virus) 2A and are inserted into the genome AI4 containing full-length transcript carrier pNDV / AI4 to obtain recombinant NDV genome full-length clone pNDV / AI4-H52AH9. The recombinant virus rNDV-H52AH9 is obtained by reverse genetic operation, has higher reproductive titer on embryonated eggs, and simultaneously expresses the HA protein of H5 and H9 subtype avian influenza virus stably, and lays a foundation for the development and application of multi-joint recombinant live vector vaccine of AI and ND.

Description

technical field [0001] The invention relates to the application of reverse genetic technology to rescue a recombinant vaccine strain rNDV-H that uses Newcastle disease virus as a carrier to co-express H5 and H9 subtype avian influenza virus hemagglutinin proteins 5 2AH 9 , for the development of vaccines. Background technique [0002] Reverse genetics manipulation technique (Reverse genetics manipulation technique) refers to the construction of infectious molecular clones of RNA viruses in vitro, the reverse transcription of viral genome RNA into cDNA, and various in vitro artificial manipulations on the DNA molecular level. A research technique for assembling new RNA viruses from genomic cDNA and various accessory proteins[ A, Mundt E, Mebatsion T, Buchholz UJ, Mettenleiter TC. Generation of recombinant lentogenic Newcastle disease virus from cDNA. J Gen Virol 1999.80(11):2987—95.], also called "virus rescue". Since the "rescued" RNA virus is derived from cDNA clones, ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/85C12R1/93
Inventor 刘秀梵丁平云宋庆庆王晓泉刘晓文胡顺林
Owner YANGZHOU UNIV
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