PubAg

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

Author:

Zhenyu Zhang, Wei Zhao, Deshan Li, Jinlong Yang, Laszlo Zsak, Oingzhong Yu

Source:

Journal of general virology 2015 v.96 no.8 pp. 2028-2035

ISSN:

0022-1317

Subject:

gene therapy, messenger RNA, quantitative analysis, phosphoproteins, open reading frames, fluorescence, vaccines, nucleocapsid, transcription (genetics), gene expression, Avian orthoavulavirus 1, pathogenicity, genes, viruses

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, 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.

Agid:

61622

Handle:

10113/61622

https://doi.org/10.1099/vir.0.000142