Improvement of the transient expression system for production of recombinant proteins in plants

Tsuyoshi Yamamoto, Ken Hoshikawa, Kentaro Ezura, Risa Okazawa, Satoshi Fujita, Miyo Takaoka, Hugh Mason, Hiroshi Ezura, Kenji Miura

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

An efficient and high yielding expression system is required to produce recombinant proteins. Furthermore, the transient expression system can be used to identify the localization of proteins in plant cells. In this study, we demonstrated that combination of a geminiviral replication and a double terminator dramatically enhanced the transient protein expression level in plants. The GFP protein was expressed transiently in lettuce, Nicotiana benthamiana, tomatoes, eggplants, hot peppers, melons, and orchids with agroinfiltration. Compared to a single terminator, a double terminator enhanced the expression level. A heat shock protein terminator combined with an extensin terminator resulted in the highest protein expression. Transiently expressed GFP was confirmed by immunoblot analysis with anti-GFP antibodies. Quantitative analysis revealed that the geminiviral vector with a double terminator resulted in the expression of at least 3.7 mg/g fresh weight of GFP in Nicotiana benthamiana, approximately 2-fold that of the geminiviral vector with a single terminator. These results indicated that combination of the geminiviral replication and a double terminator is a useful tool for transient expression of the gene of interest in plant cells.

Original languageEnglish (US)
Article number4755
JournalScientific Reports
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2018

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Nicotiana benthamiana
recombinant proteins
production technology
protein synthesis
agroinfiltration
extensin
hot peppers
eggplants
plant proteins
melons
Orchidaceae
heat shock proteins
lettuce
quantitative analysis
tomatoes
cells
gene expression
antibodies
proteins

ASJC Scopus subject areas

  • General

Cite this

Yamamoto, T., Hoshikawa, K., Ezura, K., Okazawa, R., Fujita, S., Takaoka, M., ... Miura, K. (2018). Improvement of the transient expression system for production of recombinant proteins in plants. Scientific Reports, 8(1), [4755]. https://doi.org/10.1038/s41598-018-23024-y

Improvement of the transient expression system for production of recombinant proteins in plants. / Yamamoto, Tsuyoshi; Hoshikawa, Ken; Ezura, Kentaro; Okazawa, Risa; Fujita, Satoshi; Takaoka, Miyo; Mason, Hugh; Ezura, Hiroshi; Miura, Kenji.

In: Scientific Reports, Vol. 8, No. 1, 4755, 01.12.2018.

Research output: Contribution to journalArticle

Yamamoto, T, Hoshikawa, K, Ezura, K, Okazawa, R, Fujita, S, Takaoka, M, Mason, H, Ezura, H & Miura, K 2018, 'Improvement of the transient expression system for production of recombinant proteins in plants', Scientific Reports, vol. 8, no. 1, 4755. https://doi.org/10.1038/s41598-018-23024-y
Yamamoto, Tsuyoshi ; Hoshikawa, Ken ; Ezura, Kentaro ; Okazawa, Risa ; Fujita, Satoshi ; Takaoka, Miyo ; Mason, Hugh ; Ezura, Hiroshi ; Miura, Kenji. / Improvement of the transient expression system for production of recombinant proteins in plants. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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