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

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...

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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.]

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