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E8 启动子-HSP 终止子盒促进重组蛋白在转基因番茄果实中的高水平积累:神秘果素的案例研究。

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

机构信息

Graduate School of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki, 305-8572, Japan.

出版信息

Plant Cell Rep. 2013 Apr;32(4):529-36. doi: 10.1007/s00299-013-1384-7. Epub 2013 Jan 11.

Abstract

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.

摘要

E8 启动子-HSP 终止子表达盒是一种在成熟番茄果实中提高重组蛋白积累的有力工具。在转基因植物中,强烈的、组织特异性的转基因表达是一种理想的特征,允许生产可变的重组蛋白。表达载体是控制转基因植物中转基因和重组蛋白表达水平和部位的关键工具。E8 启动子、果实成熟特异性启动子和来自拟南芥热休克蛋白 18.2 的热休克蛋白(HSP)终止子的组合,产生了在转基因番茄中强烈的、果实特异性的重组神秘果素的积累。神秘果素基因表达由 E8 启动子和 HSP 终止子盒(E8-MIR-HSP)驱动,在成熟果实中的神秘果素浓度最高,代表每克鲜重 30-630μg 的神秘果素。含有 E8-MIR-HSP 盒的转基因番茄植物系中的神秘果素浓度最高水平约为使用组成型 35S 启动子和 NOS 终止子盒(Hiwasa-Tanase 等人,在 Plant Cell Rep 30:113-124, 2011 年)观察到的水平的四倍。这些结果表明,E8 启动子和 HSP 终止子盒的组合是一种有用的工具,可以显著增加重组蛋白在成熟果实中的积累。

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