Method for constructing recombinant Newcastle disease vaccine candidate strain VII-HNF through HN gene translocation and application

A technology of VII-HNF and Newcastle disease, applied in HN gene translocation to construct recombinant Newcastle disease vaccine candidate strain VII-HNF, and in the field of vaccine development, to achieve strong immune efficacy, broad application prospects, and enhanced immune efficacy

Active Publication Date: 2022-06-03
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Hu et al obtained gene VII attenuated strain NDV / AI4 by mutation of F protein cleavage site, but this attenuated strain still has certain virulence [Hu, Z., Hu, S., Meng, C., Wang , X., Zhu, J., and Liu, X. Generation of agenotype VII Newcastle disease virus vaccine candidate with high yield inembryonated chicken eggs. Avian Dis, 2011, 55(3): 391-7.]

Method used

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  • Method for constructing recombinant Newcastle disease vaccine candidate strain VII-HNF through HN gene translocation and application
  • Method for constructing recombinant Newcastle disease vaccine candidate strain VII-HNF through HN gene translocation and application

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Embodiment Construction

[0041] The recombinant Newcastle disease vaccine candidate strain VII-HNF, whose HN gene translocation is encoded in front of the F gene, was deposited in the China Center for Type Culture Collection (Address: Wuhan University Collection Center) on November 9, 2021, and was classified and named Newcastle disease virus VII -HNF, deposit number CCTCC NO: V202177.

[0042] Construction step 1: Construction of a full-length expression clone in which the HN gene is translocated before the F gene

[0043] Biomaterial preparation:

[0044] The vector pNDV / A14 (Hu, Z., Hu, S., Meng, C., Wang, X., Zhu, J., and Liu, X.Generation of a genotype) containing the full-length genome of the Newcastle disease virus A14 strain VII Newcastle disease virusvaccine candidate with high yield in embryonated chicken eggs. Avian Dis, 2011, 55(3): 391-7.) was constructed and preserved by the Key Open Laboratory of Animal and Poultry Infectious Diseases, Ministry of Agriculture, Yangzhou University.

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Abstract

The invention discloses a method for constructing a recombinant Newcastle disease vaccine candidate strain VII-HNF through HN gene translocation and application of the recombinant Newcastle disease vaccine candidate strain VII-HNF, and the method comprises the following steps: translocating an HN gene in an AI4 genome full-length transcription vector pNDV/AI4 to an F gene by utilizing an established reverse genetic manipulation platform of a gene VII type Newcastle disease virus attenuated strain AI4; the full-length cDNA cloned pNDV/rAI4-HNF of the recombinant Newcastle disease virus genome subjected to translocation editing of the F and HN genes is obtained. The recombinant virus VII-HNF obtained by transfection has high reproduction titer and weak virulence on a chick embryo, and can be used for manufacturing vaccines.

Description

technical field [0001] The invention relates to the application of reverse genetic technology, in particular to a method for constructing recombinant Newcastle disease vaccine candidate strain VII-HNF by translocation of HN gene, and the VII-HNF is used for vaccine development. Background technique [0002] Newcastle disease is a severe infectious disease caused by a virulent strain of Newcastle Disease Virus (NDV), which is extremely harmful to the poultry industry in the world. The genome of NDV is non-segmented, single-stranded negative-stranded RNA, and the genome structure pattern of NDV is 3′-NP-P-M-F-HN-L-5′, which in turn encodes nucleocapsid protein (NP), phosphoprotein (P ), matrix protein (M), fusion protein (F), hemagglutinin-neuraminidase protein (HN) and macromolecular polymerase protein (L) six structural proteins [Yusoff, K., and Tan, W.S.Newcastle disease virus: macromolecules and opportunities. Avian Pathol, 2001, 30(5): 439-55.]. The NP, P and L proteins...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/45C12N15/56C12N15/86A61K39/17A61P31/14C12R1/93
CPCC12N7/00C07K14/005C12N9/2402C12N15/86A61K39/12A61P31/14C12Y302/01018C12N2760/18121C12N2760/18122C12N2760/18143C12N2760/18134Y02A50/30
Inventor 胡顺林刘秀梵朱珊珊王晓泉刘晓文陈玉
Owner YANGZHOU UNIV
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