Frontiers in Bioengineering and Biotechnology (Nov 2015)

Production of hybrid chimeric PVX particles using a combination of TMV and PVX-based expression vectors

  • Christina eDickmeis,
  • Mareike Michaela Antonia Honickel,
  • Rainer eFischer,
  • Ulrich eCommandeur

DOI
https://doi.org/10.3389/fbioe.2015.00189
Journal volume & issue
Vol. 3

Abstract

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We have generated hybrid chimeric Potato virus X (PVX) particles by co-expression of different PVX coat protein fusions utilizing Tobacco mosaic virus (TMV) and PVX based expression vectors. Co-infection was achieved with a modified PVX overcoat vector displaying a fluorescent protein, and a TMV vector expressing another PVX fluorescent overcoat fusion protein. Co-expression of the PVX-CP fusions in the same cells was confirmed by epifluorescence microscopy. Labeling with specific antibodies and transmission electron microscopy (TEM) revealed chimeric particles displaying GFP and mCherry on the surface. These data were corroborated by bimolecular fluorescence complementation (BiFC). We used split-mCherry fragments as PVX coat fusions and confirmed an interaction between the split-mCherry fragments in co-infected cells. The presence of assembled split-mCherry on the surface confirmed the hybrid character of the chimeric particles.

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