title: P and M gene junction is the optimal insertion site in Newcastle disease virus vaccine vector for foreign gene expression.
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 non-coding region between the P and M genes is the optimal insertion site for foreign gene expression in the VG/GA vaccine vector.
authors.affiliation: Beijing Centre for Disease Control and Prevention, Beijing 100013, PR China.|Southeast Poultry Research Laboratory, Agricultural Research Services, United States Department of Agriculture, 934 College Station Road, Athens, GA 30605, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin, Heilongjiang 150030, PR China.|Southeast Poultry Research Laboratory, Agricultural Research Services, United States Department of Agriculture, 934 College Station Road, Athens, GA 30605, USA.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Southeast Poultry Research Laboratory, Agricultural Research Services, United States Department of Agriculture, 934 College Station Road, Athens, GA 30605, USA.
authors.forename: Laszlo
authors.initials: L
authors.lastname: Zsak
authors.affiliation: Southeast Poultry Research Laboratory, Agricultural Research Services, United States Department of Agriculture, 934 College Station Road, Athens, GA 30605, USA.
authors.forename: Qingzhong
authors.initials: Q
authors.lastname: Yu
mesh_terms: D015870:Gene Expression / Q000235:genetics*; D005814:Genes, Viral / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000235:genetics*; D009522:Newcastle disease virus / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics; D014765:Viral Vaccines / Q000235:genetics*
chemical_list: D014765:Viral Vaccines; D049452:Green Fluorescent Proteins