Uniform accumulation of recombinant miraculin protein in transgenic tomato fruit using a fruit-ripening-specific E8 promoter
The E8 promoter, a tomato fruit-ripening-specific promoter, and the CaMV 35S promoter, a constitutive promoter, were used to express the miraculin gene encoding the taste-modifying protein in tomato. The accumulation of miraculin protein and mRNA was compared among transgenic tomatoes expressing the miraculin gene driven by these promoters. Recombinant miraculin protein predominantly accumulated in transgenic tomato lines using the E8 promoter (E8-MIR) only at the red fruit stage. The accumulations were almost uniform among all fruit tissues. When the 35S promoter (35S-MIR) was used, miraculin accumulation in the exocarp was much higher than in other tissues, indicating that the miraculin accumulation pattern can be regulated by using different types of promoters. We also discuss the potential of the E8-MIR lines for practical use.
KeywordsE8 promoter Miraculin Taste-modifying protein Tomato
We thank the members of the Ezura laboratory for helpful discussions. “Micro-Tom” seeds (TOMJPF00001) were obtained from the National BioResource Project Tomato (NBRP-tomato). This research was supported by the project “Development of Fundamental Technologies for the Production of High-value Materials Using Transgenic Plants” from the Ministry of Economy, Trade, and Industry of Japan (to H.E.).
- Hiwasa-Tanase K, Nyarubona M, Hirai T, Kato K, Ichikawa T, Ezura H (2011) High-level accumulation of recombinant miraculin protein in transgenic tomatoes expressing a synthetic miraculin gene with optimized codon usage terminated by the native miraculin terminator. Plant Cell Rep 30:113–124Google Scholar
- Lewinsohn E, Schalechet F, Wilkinson J, Matsui K, Tadmor Y, Nam KH, Amar O, Lastochkin E, Larkov O, Ravid U, Hiatt W, Gepstein S, Pichersky E (2001) Enhanced levels of the aroma and flavor compound S-linalool by metabolic engineering of the terpenoid pathway in tomato fruits. Plant Physiol 127:1256–1265PubMedCrossRefGoogle Scholar
- Satoh J, Kato K, Shinmyo A (2004) The 5′-untranslated region of tobacco alcohol dehydrogenase gene functions as an effective translational enhancer in plant. J Biosci Bioeng 1:1–8Google Scholar