Double subgenomic promoter control for a target gene superexpression by a plant viral vector
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Several new deconstructed vectors based on a potexvirus genome sequence for efficient expression of heterologous proteins in plants were designed. The first obtained vector (AltMV-single), based on the Alternanthera mosaic virus (AltMV) strain MU genome, bears a typical architecture for deconstructed plant viral vectors, i.e. a triple gene block was deleted from the viral genome and the model gene of interest was placed under control of the first viral subgenomic promoter. To enhance the efficiency of expression, maintained by the AltMV-single, another vector (AltMV-double) was designed. In AltMV-double, the gene of interest was controlled by two viral subgenomic promoters located sequentially without a gap upstream of the target gene. It was found that AltMV-double provided a significantly higher level of accumulation of the target protein in plants than AltMV-single. Moreover, our data clearly show the requirement of the presence and functioning of both the subgenomic promoters for demonstrated high level of target protein expression by AltMV-double. Taken together, our results describe an additional possible way to enhance the efficiency of transient protein expression maintained in plants by a plant viral vector.
Key wordsAlternanthera mosaic virus viral vector protein overexpression subgenomic promoter
Alternanthera mosaic virus
fresh leaf tissue
human granulocyte colony-stimulating factor
open reading frame
triple gene block
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- 9.Tilsner, J., Linnik, O., Wright, K. M., Bell, K., Roberts, A. G., Lacomme, C., Santa Cruz, S., and Oparka, K. J. (2012) The TGB1 movement protein of potato virus X reorganizes actin and endomembranes into the X-body, a viral replication factory, Plant Physiol., 158, 1359–1370.PubMedCentralCrossRefPubMedGoogle Scholar
- 13.Ivanov, P. A., Mukhamedzhanova, A. A., Smirnov, A. A., Rodionova, N. P., Karpova, O. V., and Atabekov, J. G. (2010) The complete nucleotide sequence of Alternanthera mosaic virus infecting Portulaca grandiflora represents a new strain distinct from phlox isolates, Virus Genes, 42, 268–271.CrossRefPubMedGoogle Scholar
- 14.Lim, H. S., Vaira, A. M., Reinsel, M. D., Bae, H., Bailey, B. A., Domier, L. L., and Hammond, J. (2010) Pathogenicity of Alternanthera mosaic virus is affected by determinants in RNA-dependent RNA polymerase and by reduced efficacy of silencing suppression in a movement-competent TGB1, J. Gen. Virol., 91, 277–287.CrossRefPubMedGoogle Scholar
- 16.Zvereva, A. S., Petrovskaya, L. E., Rodina, A. V., Frolova, O. Y., Ivanov, P. A., Shingarova, L. N., Komarova, T. V., Dorokhov, Y. L., Dolgikh, D. A., Kirpichnikov, M. P., and Atabekov, J. G. (2009) Production of biologically active human myelocytokines in plants, Biochemistry (Moscow), 74, 1187–1194.PubMedGoogle Scholar