Taniguchi Naohiro, Murakami Hiroshi
PeptiDream Inc., 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8505, Japan.
Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Methods Mol Biol. 2017;1498:339-347. doi: 10.1007/978-1-4939-6472-7_22.
Constructing protein-coding genes with desired mutations is a basic step for protein engineering. Herein, we describe a multiple site-directed and saturation mutagenesis method, termed MUPAC. This method has been used to introduce multiple site-directed mutations in the green fluorescent protein gene and in the moloney murine leukemia virus reverse transcriptase gene. Moreover, this method was also successfully used to introduce randomized codons at five desired positions in the green fluorescent protein gene, and for simple DNA assembly for cloning.
构建具有所需突变的蛋白质编码基因是蛋白质工程的基本步骤。在此，我们描述了一种称为MUPAC的多位点定向和饱和诱变方法。该方法已用于在绿色荧光蛋白基因和莫洛尼鼠白血病病毒逆转录酶基因中引入多位点定向突变。此外，该方法还成功用于在绿色荧光蛋白基因的五个所需位置引入随机密码子，并用于克隆的简单DNA组装。
