Extremely High-Level and Rapid Transient Protein Production in Plants without the Use of Viral Replication

Sainsbury, Frank and Lomonossoff, George P. (2008) Extremely High-Level and Rapid Transient Protein Production in Plants without the Use of Viral Replication. Plant Physiology, 148 3: 1212-1218. doi:10.1104/pp.108.126284


Author Sainsbury, Frank
Lomonossoff, George P.
Title Extremely High-Level and Rapid Transient Protein Production in Plants without the Use of Viral Replication
Journal name Plant Physiology   Check publisher's open access policy
ISSN 0032-0889
1532-2548
Publication date 2008-11-01
Year available 2008
Sub-type Article (original research)
DOI 10.1104/pp.108.126284
Volume 148
Issue 3
Start page 1212
End page 1218
Total pages 7
Place of publication Rockville, MD United States
Publisher American Society of Plant Biologists
Collection year 2008
Language eng
Formatted abstract
Plant-based overexpression of heterologous proteins has attracted much interest and development in recent years. To date, the most efficient vectors have been based on RNA virus-derived replicons. A system based on a disabled version of cowpea mosaic virus RNA-2 has been developed, which overcomes limitations on insert size and introduces biocontainment. This system involves positioning a gene of interest between the 5′ leader sequence and 3′ untranslated region (UTR) of RNA-2, thereby emulating a presumably stable mRNA for efficient translation. Thus far, the sequence of the 5′ UTR has been preserved to maintain the ability of the modified RNA-2 to be replicated by RNA-1. However, high-level expression may be achieved in the absence of RNA-1-derived replication functions using Agrobacterium-mediated transient transformation. To investigate those features of the 5′ UTR necessary for efficient expression, we have addressed the role of two AUG codons found within the 5′ leader sequence upstream of the main initiation start site. Deletion of an in-frame start codon upstream of the main translation initiation site led to a massive increase in foreign protein accumulation. By 6 d postinfiltration, a number of unrelated proteins, including a full-size IgG and a self-assembling virus-like particle, were expressed to >10% and 20% of total extractable protein, respectively. Thus, this system provides an ideal vehicle for high-level expression that does not rely on viral replication of transcripts. 
Keyword Heterologous proteins
Translation
Antibodies
Systems
Vector
Transformation
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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