Ge Xumeng, Xu Jianfeng
Arkansas Biosciences Institute, Arkansas State University, Jonesboro, AR, USA.
Methods Mol Biol. 2012;824:565-78. doi: 10.1007/978-1-61779-433-9_30.
Cell-free protein synthesis (CFPS) has major advantages over traditional cell-based methods in the capability of high-throughput protein synthesis and special protein production. During recent decades, CFPS has become an alternative protein production platform for both fundamental and applied purposes. Using Renilla luciferase as model protein, we describe a typical process of CFPS in wheat germ extract system, including wheat germ extract preparation, expression vector construction, in vitro protein synthesis (transcription/translation), and target protein assay.
无细胞蛋白质合成(CFPS)在高通量蛋白质合成能力和特殊蛋白质生产方面比传统的基于细胞的方法具有主要优势。在最近几十年中,CFPS已成为用于基础和应用目的的替代蛋白质生产平台。以海肾荧光素酶作为模型蛋白,我们描述了小麦胚芽提取物系统中CFPS的典型过程,包括小麦胚芽提取物的制备、表达载体的构建、体外蛋白质合成(转录/翻译)以及目标蛋白的检测。