Expression of multiple proteins using full-length and deleted versions of cowpea mosaic virus RNA-2

Sainsbury, Frank, Lavoie, Pierre-Olivier, D'Aoust, Marc-Andre, Vezina, Louis-Philippe and Lomonossoff, George P. (2008) Expression of multiple proteins using full-length and deleted versions of cowpea mosaic virus RNA-2. Plant Biotechnology Journal, 6 1: 82-92. doi:10.1111/j.1467-7652.2007.00303.x

Author Sainsbury, Frank
Lavoie, Pierre-Olivier
D'Aoust, Marc-Andre
Vezina, Louis-Philippe
Lomonossoff, George P.
Title Expression of multiple proteins using full-length and deleted versions of cowpea mosaic virus RNA-2
Journal name Plant Biotechnology Journal   Check publisher's open access policy
ISSN 1467-7644
Publication date 2008-01
Sub-type Article (original research)
DOI 10.1111/j.1467-7652.2007.00303.x
Open Access Status
Volume 6
Issue 1
Start page 82
End page 92
Total pages 11
Place of publication Oxford, United Kingdom
Publisher Wiley-Blackwell Publishing
Language eng
Formatted abstract
The use of multiple copies of vectors based on either full-length or deleted versions of cowpea mosaic virus RNA-2 for the production of heteromeric proteins in plants was investigated. Co-infiltration of two full-length RNA-2 constructs containing different marker genes into Nicotiana benthamiana in the presence of RNA-1 showed that the two foreign proteins were efficiently expressed within the same cell in inoculated tissue. Furthermore, the proteins were co-localized to the same subcellular compartments, an essential prerequisite for heteromer formation. However, segregation of two separate RNA-2 molecules, and therefore expression of the two proteins, was observed on systemic spread of the recombinant viruses. Thus, efficient assembly of heteromeric proteins is likely to occur only in inoculated tissue. To determine the optimum approach for expression in inoculated tissue, the heavy and light chains of the blood group-typing immunoglobulin G (IgG) C5-1 were inserted into full-length and deleted versions of RNA-2, and the constructs were agroinfiltrated in the presence of RNA-1. The results obtained showed that full-size IgG molecules accumulated using both approaches, but that the levels were significantly higher when deleted RNA-2 vectors were used. The levels were also greatly enhanced by the inclusion of an endoplasmic reticulum retention signal at the C-terminus of the heavy chain. As the potential benefit of using full-length RNA-2 constructs, the ability to spread systemically, appears to be irrelevant to the production of heteromeric proteins, the use of deleted versions of RNA-2 is clearly advantageous, particularly as they offer the benefit of biocontainment.
Keyword Cowpea mosaic virus
Deleted RNA-2
Fluorescent protein
Molecular farming
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
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