Incorporation of regulatory DNA elements within a viral vector improves recombinant protein expression in plants

Coates, Ryan J., Scofield, Simon ORCID: https://orcid.org/0000-0001-5466-314X and Young, Mark T. ORCID: https://orcid.org/0000-0002-9615-9002 2024. Incorporation of regulatory DNA elements within a viral vector improves recombinant protein expression in plants. Scientific Reports 14 (1) , 28865. 10.1038/s41598-024-80444-9

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Abstract

Plants have significant potential as recombinant protein expression chassis, as they can produce complex post-translationally modified proteins that are unobtainable using prokaryotic production systems, with almost limitless scalability and substantially reduced costs relative to eukaryotic cell cultures. Transient protein expression reduces the time taken between transformation and recombinant protein extraction and purification, however low protein yields relative to conventional stable expression systems remain a major obstacle. Here, we have assessed the effectiveness of combining several established genetic components, including a promoter, 5’ UTR, 3’ UTR, double terminator, and matrix attachment region, to modify the TMV-based pJL-TRBO expression vector for improved recombinant protein expression in plants. Using enhanced green fluorescent protein (eGFP) as a reporter, we quantified expression using fluorescence imaging in planta together with SDS-PAGE and western blotting and showed that our optimum construct resulted in a significant increase relative to pJL-TRBO-eGFP. This increase was exclusively due to the presence of the additional 5’ UTR. We anticipate that our expression constructs will be a useful tool for high-yield plant recombinant protein production and may serve as a template for further improvements.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	Biosciences
Additional Information:	License information from Publisher: LICENSE 1: URL: http://creativecommons.org/licenses/by/4.0/, Type: open-access
Publisher:	Nature Research
Funders:	BBSRC
Date of First Compliant Deposit:	22 November 2024
Date of Acceptance:	19 November 2024
Last Modified:	04 Dec 2024 12:42
URI:	https://orca.cardiff.ac.uk/id/eprint/174236

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