An E8 promoter–HSP terminator cassette promotes the high-level accumulation of recombinant protein predominantly in transgenic tomato fruits: a case study of miraculin

Plant Cell Reports volume 32, pages529–536(2013)Cite this article

Abstract

Key message

The E8 promoter–HSP terminator expression cassette is a powerful tool for increasing the accumulation of recombinant protein in a ripening tomato fruit.

Abstract

Strong, tissue-specific transgene expression is a desirable feature in transgenic plants to allow the production of variable recombinant proteins. The expression vector is a key tool to control the expression level and site of transgene and recombinant protein expression in transgenic plants. The combination of the E8 promoter, a fruit-ripening specific promoter, and a heat shock protein (HSP) terminator, derived from heat shock protein 18.2 of Arabidopsis thaliana, produces the strong and fruit-specific accumulation of recombinant miraculin in transgenic tomato. Miraculin gene expression was driven by an E8 promoter and HSP terminator cassette (E8–MIR–HSP) in transgenic tomato plants, and the miraculin concentration was the highest in the ripening fruits, representing 30–630 μg miraculin of the gram fresh weight. The highest level of miraculin concentration among the transgenic tomato plant lines containing the E8–MIR–HSP cassette was approximately four times higher than those observed in a previous study using a constitutive 35S promoter and NOS terminator cassette (Hiwasa-Tanase et al. in Plant Cell Rep 30:113–124, 2011). These results demonstrate that the combination of the E8 promoter and HSP terminator cassette is a useful tool to increase markedly the accumulation of recombinant proteins in a ripening fruit-specific manner.

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Acknowledgments

We thank the members of the Ezura laboratory for their helpful discussions. This study was supported by the project ‘‘Development of Fundamental Technologies for the Production of High-value Materials Using Transgenic Plants’’ by the Ministry of Economy, Trade, and Industry of Japan (to H.E.). The Micro-Tom seeds (TOMJPF00001) were provided by the National BioResource Project (NBRP) of the MEXT, Japan.

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Affiliations

  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki, 305-8572, Japan

    Natsuko Kurokawa, Tadayoshi Hirai, Mariko Takayama, Kyoko Hiwasa-Tanase & Hiroshi Ezura

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  1. Natsuko Kurokawa
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  2. Tadayoshi Hirai
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  3. Mariko Takayama
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  4. Kyoko Hiwasa-Tanase
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  5. Hiroshi Ezura
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Correspondence to Hiroshi Ezura.

Additional information

Communicated by H. Ebinuma.

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Kurokawa, N., Hirai, T., Takayama, M. et al. An E8 promoter–HSP terminator cassette promotes the high-level accumulation of recombinant protein predominantly in transgenic tomato fruits: a case study of miraculin. Plant Cell Rep 32, 529–536 (2013). https://doi.org/10.1007/s00299-013-1384-7

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Keywords

  • Expression vector
  • E8 promoter
  • HSP terminator
  • Miraculin
  • Transgenic tomato plants