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

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. Strong, tissue-specific transgene expression is a desirable feature in transgenic plants to allow the production of variable recombin...

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Bibliographic Details
Published in:Plant cell reports 2013-04, Vol.32 (4), p.529-536
Main Authors: Kurokawa, Natsuko, Hirai, Tadayoshi, Takayama, Mariko, Hiwasa-Tanase, Kyoko, Ezura, Hiroshi
Format: Article
Language:English
Subjects:
Arabidopsis
Arabidopsis Proteins - genetics
Arabidopsis thaliana
Biomedical and Life Sciences
Biotechnology
case studies
Cell Biology
Fruit - genetics
Fruit - metabolism
fruits
gene expression
Gene Expression Regulation, Plant
Glycoproteins - biosynthesis
Glycoproteins - genetics
heat shock proteins
Heat-Shock Proteins - genetics
Life Sciences
Lycopersicon esculentum
Lycopersicon esculentum - genetics
Lycopersicon esculentum - metabolism
Original Paper
Plant Biochemistry
Plant Sciences
Plants, Genetically Modified - genetics
Plants, Genetically Modified - metabolism
Promoter Regions, Genetic
protein synthesis
recombinant proteins
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Terminator Regions, Genetic
tomatoes
transgenic plants
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Summary: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. 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.
ISSN:0721-7714
1432-203X
DOI:10.1007/s00299-013-1384-7