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P and M gene junction is the optimal insertion site in Newcastle disease virus vaccine vector for foreign gene expression

Wei Zhao, Zhenyu Zhang, Laszlo Zsak, Oingzhong Yu
Journal of general virology 2015 v.96 no.Pt. 1 pp. 40-45
gene therapy, green fluorescent protein, messenger RNA, fluorescence, vaccines, gene expression, Avian orthoavulavirus 1, viruses, genes, intergenic DNA, pathotypes
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 39 end and gradually decreased as the gene was inserted closer to the 59 end. Measurement of the GFP fluorescence intensity in recombinant virus-infected cells demonstrated that the non-coding region between the P and M genes is the optimal insertion site for foreign gene expression in the VG/GA vaccine vector.