title: Development of a Newcastle disease virus vector expressing a foreign gene through an internal ribosomal entry site provides direct proof for a sequential transcription mechanism.
abstract: In the present study, we developed a novel approach for foreign gene expression by Newcastle disease virus (NDV) from a second ORF through an internal ribosomal entry site (IRES). Six NDV LaSota strain-based recombinant viruses vectoring the IRES and a red fluorescence protein (RFP) gene behind the nucleocapsid (NP), phosphoprotein (P), matrix (M), fusion (F), haemagglutinin-neuraminidase (HN) or large polymerase (L) gene ORF were generated using reverse genetics technology. The insertion of the second ORF slightly attenuated virus pathogenicity, but did not affect ability of the virus to grow. Quantitative measurements of RFP expression in virus-infected DF-1 cells revealed that the abundance of viral mRNAs and red fluorescence intensity were positively correlated with the gene order of NDV, 3'-NP-P-M-F-HN-L-5', proving the sequential transcription mechanism for NDV. The results herein suggest that the level of foreign gene expression could be regulated by selecting the second ORF insertion site to maximize the efficacy of vaccine and gene therapy.
authors: College of Life Sciences, Northeast Agricultural University, Harbin, Heilongjiang 150030, PR China.|United States Department of Agriculture, U.S. National Poultry Research Center, Agricultural Research Services, 934 College Station Road, Athens, GA, 30605, USA.
authors: Zhenyu
authors: Z
authors: Zhang
authors: United States Department of Agriculture, U.S. National Poultry Research Center, Agricultural Research Services, 934 College Station Road, Athens, GA, 30605, USA.|Beijing Centre for Disease Control and Prevention, Beijing 100013, PR China.
authors: Wei
authors: W
authors: Zhao
authors: College of Life Sciences, Northeast Agricultural University, Harbin, Heilongjiang 150030, PR China.
authors: Deshan
authors: D
authors: Li
authors: United States Department of Agriculture, U.S. National Poultry Research Center, Agricultural Research Services, 934 College Station Road, Athens, GA, 30605, USA.|Chongqing Academy of Animal Sciences, Chongqing 402460, PR China.
authors: Jinlong
authors: J
authors: Yang
authors: United States Department of Agriculture, U.S. National Poultry Research Center, Agricultural Research Services, 934 College Station Road, Athens, GA, 30605, USA.
authors: Laszlo
authors: L
authors: Zsak
authors: United States Department of Agriculture, U.S. National Poultry Research Center, Agricultural Research Services, 934 College Station Road, Athens, GA, 30605, USA.
authors: Qingzhong
authors: Q
authors: Yu
mesh_terms: D000818:Animals; D002645:Chickens; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D009521:Newcastle Disease; D009522:Newcastle disease virus / Q000235:genetics* / Q000378:metabolism; D011201:Poultry Diseases; D059386:Reverse Genetics / Q000379:methods*; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014765:Viral Vaccines / Q000235:genetics / Q000378:metabolism
chemical_list: D014764:Viral Proteins; D014765:Viral Vaccines