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Biochemistry (Moscow)

, Volume 80, Issue 8, pp 1039–1046 | Cite as

Double subgenomic promoter control for a target gene superexpression by a plant viral vector

  • E. V. PutlyaevEmail author
  • A. A. Smirnov
  • O. V. Karpova
  • J. G. Atabekov
Article

Abstract

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 words

Alternanthera mosaic virus viral vector protein overexpression subgenomic promoter 

Abbreviations

AltMV

Alternanthera mosaic virus

CP

capsid protein

d.p.i.

days post-injection

f.l.t.

fresh leaf tissue

hG-CSF

human granulocyte colony-stimulating factor

ORF

open reading frame

sgp

subgenomic promoter

sgRNA

subgenomic RNA

TGB

triple gene block

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Copyright information

© Pleiades Publishing, Ltd. 2015

Authors and Affiliations

  • E. V. Putlyaev
    • 1
    Email author
  • A. A. Smirnov
    • 1
  • O. V. Karpova
    • 1
  • J. G. Atabekov
    • 1
    • 2
  1. 1.Faculty of BiologyLomonosov Moscow State UniversityMoscowRussia
  2. 2.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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