Eukaryotic in vitro translation systems require large numbers of protein and RNA components and thereby rely on the use of cell extracts. Here we established a new in vitro translation system based on rice callus extract (RCE). We confirmed that RCE maintains its initial activity even after five freeze-thaw cycles and that the optimum temperature for translation is around 20°C. We demonstrated that the RCE system allows the synthesis of hERG, a large membrane protein, in the presence of liposomes. We also showed that the introduction of a bicistronic mRNA based on 2A peptide to RCE allowed the production of two distinct proteins from a single mRNA. Our new method thus facilitates laboratory-scale production of cell extracts, making it a useful tool for the in vitro synthesis of proteins for biochemical studies.

真核体外翻译系统需要大量的蛋白质和RNA成分，因此依赖于细胞提取物的使用。在这里，我们建立了一个基于水稻愈伤组织提取物（RCE）的新的体外翻译系统。我们确认，即使经过五个冻融循环，RCE仍能保持其初始活性，并且翻译的最佳温度约为20°C。我们证明了RCE系统可以在脂质体存在下合成hERG，一种大的膜蛋白。我们还表明，将基于2A肽的双顺反子mRNA引入RCE可以从单个mRNA中产生两种不同的蛋白质。因此，我们的新方法有助于实验室规模的细胞提取物生产，使其成为体外有用的工具 用于生化研究的蛋白质合成。