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Adaptive mutation in nuclear export protein allows stable transgene expression in a chimaeric influenza A virus vector
- Source :
- The Journal of general virology. 95(Pt 2)
- Publication Year :
- 2013
-
Abstract
- The development of influenza virus vectors with long insertions of foreign sequences remains difficult due to the small size and instable nature of the virus. Here, we used the influenza virus inherent property of self-optimization to generate a vector stably expressing long transgenes from the NS1 protein ORF. This was achieved by continuous selection of bright fluorescent plaques of a GFP-expressing vector during multiple passages in mouse B16f1 cells. The newly generated vector acquired stability in IFN-competent cell lines and in vivo in murine lungs. Although improved vector fitness was associated with the appearance of four coding mutations in the polymerase (PB2), haemagglutinin and non-structural (NS) segments, the stability of the transgene expression was dependent primarily on the single mutation Q20R in the nuclear export protein (NEP). Importantly, a longer insert, such as a cassette of 1299 nt encoding two Mycobacterium tuberculosis Esat6 and Ag85A proteins, could substitute for the GFP transgene. Thus, the inherent property of the influenza virus to adapt can also be used to adjust a vector backbone to give stable expression of long transgenes.
- Subjects :
- Transgene
Recombinant Fusion Proteins
Genetic Vectors
Green Fluorescent Proteins
Active Transport, Cell Nucleus
Gene Expression
Biology
Viral Nonstructural Proteins
medicine.disease_cause
Virus Replication
Virus
Insert (molecular biology)
Green fluorescent protein
Cell Line
Mice
Genes, Reporter
Virology
Gene expression
Influenza A virus
medicine
Animals
Vector (molecular biology)
Nuclear export signal
Viral Load
Recombinant Proteins
Subjects
Details
- ISSN :
- 14652099
- Volume :
- 95
- Issue :
- Pt 2
- Database :
- OpenAIRE
- Journal :
- The Journal of general virology
- Accession number :
- edsair.doi.dedup.....0b2d34841471c977ca3d79ea5e9903a8