Contreras R, Cheroutre H, Degrave W, Fiers W
Nucleic Acids Res. 1982 Oct 25;10(20):6353-62. doi: 10.1093/nar/10.20.6353.
A simple and efficient method for direct in vitro synthesis of capped transcripts of cloned eukaryotic genes is described. As an example capped transcripts were made from a plasmid containing the human fibroblast interferon gene cloned under the control of a prokaryotic promoter. These transcripts were translated in vivo in Xenopus laevis oocytes and in vitro in reticulocyte and in wheat germ cell-free protein synthesizing systems.
本文描述了一种简单高效的直接体外合成克隆真核基因加帽转录本的方法。作为示例,从一个含有在原核启动子控制下克隆的人成纤维细胞干扰素基因的质粒制备了加帽转录本。这些转录本在非洲爪蟾卵母细胞中进行体内翻译,并在网织红细胞和麦胚无细胞蛋白质合成系统中进行体外翻译。