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

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...

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Bibliographic Details
Published in:Journal of general virology 2015-08, Vol.96 (8), p.2028-2035
Main Authors: Zhang, Zhenyu, Zhao, Wei, Li, Deshan, Yang, Jinlong, Zsak, Laszlo, Yu, Oingzhong
Format: Article
Language:English
Subjects:
Animals
Chickens
fluorescence
Gene Expression
Gene Expression Regulation, Viral
gene therapy
genes
Genetic Vectors - genetics
Genetic Vectors - metabolism
messenger RNA
Newcastle Disease
Newcastle disease virus
Newcastle disease virus - genetics
Newcastle disease virus - metabolism
nucleocapsid
open reading frames
pathogenicity
phosphoproteins
Poultry Diseases
quantitative analysis
Reverse Genetics - methods
Ribosomes - genetics
Ribosomes - metabolism
transcription (genetics)
Transcription, Genetic
vaccines
Viral Proteins - genetics
Viral Proteins - metabolism
Viral Vaccines - genetics
Viral Vaccines - metabolism
viruses
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Summary: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, 39-NP-P-M-F-HNL- 59, 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.
ISSN:0022-1317
1465-2099
DOI:10.1099/vir.0.000142