TY - JOUR
T1 - The HSP terminator of Arabidopsis thaliana induces a high level of miraculin accumulation in transgenic tomatoes
AU - Hirai, Tadayoshi
AU - Kurokawa, Natsuko
AU - Duhita, Narendra
AU - Hiwasa-Tanase, Kyoko
AU - Kato, Kazuhisa
AU - Kato, Ko
AU - Ezura, Hiroshi
PY - 2011/9/28
Y1 - 2011/9/28
N2 - High-level accumulation of the target recombinant protein is a significant issue in heterologous protein expression using transgenic plants. Miraculin, a taste-modifying protein, was accumulated in transgenic tomatoes using an expression cassette in which the miraculin gene was expressed by the cauliflower mosaic virus (CaMV) 35S promoter and the heat shock protein (HSP) terminator (MIR-HSP). The HSP terminator was derived from heat shock protein 18.2 in Arabidopsis thaliana. Using this HSP-containing cassette, the miraculin concentration in T 0 transgenic tomato lines was 1.4-13.9% of the total soluble protein (TSP), and that in the T 1 transgenic tomato line homozygous for the miraculin gene reached 17.1% of the TSP. The accumulation level of the target protein was comparable to levels observed with chloroplast transformation. The high-level accumulation of miraculin in T 0 transgenic tomato lines achieved by the HSP terminator was maintained in the successive T 1 generation, demonstrating the genetic stability of this accumulation system.
AB - High-level accumulation of the target recombinant protein is a significant issue in heterologous protein expression using transgenic plants. Miraculin, a taste-modifying protein, was accumulated in transgenic tomatoes using an expression cassette in which the miraculin gene was expressed by the cauliflower mosaic virus (CaMV) 35S promoter and the heat shock protein (HSP) terminator (MIR-HSP). The HSP terminator was derived from heat shock protein 18.2 in Arabidopsis thaliana. Using this HSP-containing cassette, the miraculin concentration in T 0 transgenic tomato lines was 1.4-13.9% of the total soluble protein (TSP), and that in the T 1 transgenic tomato line homozygous for the miraculin gene reached 17.1% of the TSP. The accumulation level of the target protein was comparable to levels observed with chloroplast transformation. The high-level accumulation of miraculin in T 0 transgenic tomato lines achieved by the HSP terminator was maintained in the successive T 1 generation, demonstrating the genetic stability of this accumulation system.
KW - CaMV 35S promoter
KW - high-level expression
KW - HSP terminator
KW - miraculin
KW - transgenic tomato plants
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U2 - 10.1021/jf202501e
DO - 10.1021/jf202501e
M3 - Article
C2 - 21861502
AN - SCOPUS:80053125767
SN - 0021-8561
VL - 59
SP - 9942
EP - 9949
JO - Journal of Agricultural and Food Chemistry
JF - Journal of Agricultural and Food Chemistry
IS - 18
ER -