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Manually-combined Newcastle diseases virus F gene and recombining expression vector and application thereof

A technology of chicken Newcastle disease virus and expression vector, applied in the field of cDNA, to achieve the effect of preventing proliferation

Inactive Publication Date: 2010-01-20
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The technical problem to be solved in the present invention at first is to overcome the deficiencies in the prior art, and the Newcastle disease virus F 48 E. 9 Molecular transformation and modification of the F gene of the newcastle disease virus to provide an artificial F gene of Newcastle disease virus that is more suitable for high-efficiency expression in chicken somatic cells

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  • Manually-combined Newcastle diseases virus F gene and recombining expression vector and application thereof
  • Manually-combined Newcastle diseases virus F gene and recombining expression vector and application thereof
  • Manually-combined Newcastle diseases virus F gene and recombining expression vector and application thereof

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Embodiment Construction

[0047] The present invention will be further described below in conjunction with specific embodiments, and the advantages and characteristics of the present invention will become clearer along with the description. However, these embodiments are only exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that the details and forms of the technical solutions of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the protection scope of the present invention.

[0048] 1 Materials and methods

[0049] 1.1 Materials

[0050] 1.1.1 Plasmids, Bacteria, Viruses and Cells

[0051] Plasmid pVAX1 was purchased from Invitrogen (Catalog Number: V26020); NDV F 48 E. 9 Strains of virus were purchased from China Veterinary Drug Control Institute (species preservation number: AV1611); DH5α hos...

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Abstract

The invention discloses a manually-combined Newcastle diseases virus F gene and a recombining expression vector and an application thereof. Codons of NDVF gene OFR are all replaced into chicken bias codons, EcoR V restriction sites and Kozak sequences are added upstream, Xbal I restriction sites and TGA stop codons are added downstream, and downstream bases A of the stop codons are changed into G, obtained genes (SEQ, ID NO: 2) can be efficiently expressed in eukaryotic cells, and the expression efficiency of the obtained genes is higher than that of wild type genes. Expressed genes have immunogenicity and biologic activity of natural protein of Newcastle diseases virus, can stimulate chicken to generate body fluid stronger immunologic response and stronger cell immunologic response than the wild F gene DNA vaccine so as to enable immunized chicken to obtain 100 percent of resistance to fatal attack of high-pathogenicity Newcastle strains, and can effectively inhibit the propagation of the virus in the chicken.

Description

technical field [0001] The invention relates to a cDNA, in particular to an artificially transformed chicken Newcastle disease virus E 48 f 9 The strain F gene and the recombinant eukaryotic expression vector containing the F gene, the invention also relates to their application as a DNA vaccine in the prevention or treatment of chicken Newcastle disease, which belongs to the field of prevention and control of chicken Newcastle disease. Background technique [0002] Newcastle disease virus (NDV) belongs to the genus Mumps virus in the Paramyxoviridae family. It is a highly pathogenic virus that is harmful to poultry and is recognized as one of the two important poultry diseases in the world. The case fatality rate of this disease is extremely high, and the highly virulent virus strain can often cause 100% infection of susceptible flocks, and cause 100% death, has spread all over the world. At present, traditional vaccines such as oil adjuvant inactivated vaccines or attenu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/45C12N15/63C12N5/10A61K48/00A61K39/17A61P31/14C12R1/93
Inventor 王云峰石星明王玫童光志曾伟伟
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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