The HSP Terminator of Arabidopsis thaliana Induces a High Level of Miraculin Accumulation in Transgenic Tomatoes
The HSP Terminator of Arabidopsis thaliana Induces a High Level of Miraculin Accumulation in Transgenic Tomatoes
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DOI：
10.1021/jf202501e
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发表时间：
2011-09-28
影响因子：
6.1
通讯作者：
Ezura, Hiroshi
中科院分区：
文献类型：
--
作者：
Hirai, Tadayoshi;Kurokawa, Natsuko;Ezura, Hiroshi
目标重组蛋白的高水平积累是使用转基因植物表达异源蛋白的一个重要问题。 Miraculin 是一种味道调节蛋白，使用表达盒在转基因番茄中积累，其中 Miraculin 基因由花椰菜花叶病毒 (CaMV) 35S 启动子和热休克蛋白 (HSP) 终止子 (MIR-HSP) 表达。 HSP 终止子源自拟南芥中的热休克蛋白 18.2。使用该含有HSP的盒，在To转基因番茄品系中，奇迹蛋白浓度为总可溶性蛋白(TSP)的1.4-13.9％，而在奇迹蛋白基因纯合的T-1转基因番茄品系中，奇迹蛋白浓度达到TSP的17.1％。目标蛋白的积累水平与叶绿体转化观察到的水平相当。通过HSP终止子实现的To转基因番茄品系中奇迹蛋白的高水平积累在连续的T-1代中得以维持，证明了该积累系统的遗传稳定性。
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 To 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 To transgenic tomato lines achieved by the HSP terminator was maintained in the successive T-1 generation, demonstrating the genetic stability of this accumulation system.