|ZHAO, WEI - Beijing Center For Diseases Prevention And Control|
|ZHANG, ZHENYU - Northeast Agricultural University|
Submitted to: Journal of General Virology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/28/2014
Publication Date: 1/2/2015
Publication URL: http://handle.nal.usda.gov/10113/60583
Citation: Zhao, W., Zhang, Z., Zsak, L., Yu, Q. 2015. P and M gene junction is the optimal insertion site in Newcastle disease virus vaccine vector for foreign gene expression. Journal of General Virology. 96:40-45.
Interpretive Summary: Poultry enteric viral diseases are responsible for considerable economic losses to the poultry industry worldwide. Unfortunately, many of the enteric viruses are uncultivatable, which restricts conventional vaccine development. Therefore, there is an urgent need to discover and develop novel vaccine strategies. In the present study, we developed an enterotropic strain of Newcastle disease virus (NDV) as a vaccine vector and systemically evaluated the effects of the insertion site at every intergenic region of the NDV genome on foreign gene expression. For the first time we have experimentally proved the NDV sequential transcription theory and identified the noncoding region between the P and M genes is the optimal insertion site for foreign genes expression. This novel vaccine vector can be used to express enteric virus antigens in the gut of young birds to prevent Newcastle disease and the targeted enteric viral diseases of poultry.
Technical Abstract: Newcastle disease virus (NDV) has been developed as a vector for vaccine and gene therapy purposes. However, the optimal insertion site for foreign gene expression remained to be determined. In the present study, we inserted the green fluorescence protein (GFP) gene into five different intergenic regions of the enterotropic NDV VG/GA vaccine strain using reverse genetics technology. The rescued recombinant viruses retained lentogenic pathotype and displayed delayed growth dynamics, particularly when the GFP gene was inserted between the NP and P genes of the virus. The GFP mRNA level was most abundant when the gene was inserted closer to the 3’ end and gradually decreased as the gene was inserted closer to the 5’ end. Measurement of the GFP fluorescence intensity in recombinant virus-infected cells demonstrated that the noncoding region between the P and M genes is the optimal insertion site for foreign gene expression in the VG/GA vaccine vector.