Graduate School of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305-8572, Japan.
J Agric Food Chem. 2011 Sep 28;59(18):9942-9. doi: 10.1021/jf202501e. Epub 2011 Sep 6.
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 T0 transgenic tomato lines was 1.4-13.9% of the total soluble protein (TSP), and that in the T1 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 T0 transgenic tomato lines achieved by the HSP terminator was maintained in the successive T1 generation, demonstrating the genetic stability of this accumulation system.
高水平积累目标重组蛋白是利用转基因植物进行异源蛋白表达的一个重大问题。甜味修饰蛋白神秘果素(miraculin)在转基因番茄中积累,使用的表达盒中,神秘果素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)驱动表达。HSP 终止子源自拟南芥的热休克蛋白 18.2。使用这个 HSP 包含的盒,T0 代转基因番茄系中神秘果素的浓度为总可溶性蛋白(TSP)的 1.4-13.9%,而在神奇果素基因纯合的 T1 代转基因番茄系中达到 TSP 的 17.1%。目标蛋白的积累水平与叶绿体转化中观察到的水平相当。通过 HSP 终止子在 T0 代转基因番茄系中实现的神秘果素高水平积累在随后的 T1 代中得以维持,表明该积累系统具有遗传稳定性。