Olins P O, Devine C S, Rangwala S H, Kavka K S
Biological Sciences Department, Monsanto Company, St. Louis, MO 63198.
Gene. 1988 Dec 15;73(1):227-35. doi: 10.1016/0378-1119(88)90329-0.
Expression of foreign genes in Escherichia coli requires the juxtaposition of prokaryotic transcription and translation elements with a coding region for the foreign gene. Commonly, this results in only modest expression of the foreign gene product. Here we describe a novel ribosome-binding site (RBS; phage T7 'gene 10 leader') which is able to drive the translation of several foreign genes. This RBS dramatically enhanced the translation efficiency of all the genes we have tested to date, and was particularly effective for foreign genes. The enhanced expression was often more than 40-fold greater than that obtained using a 'consensus' RBS. A general plasmid vector has been constructed, incorporating the T7 gene 10 leader sequence, which allows the facile expression of important gene products. In this report we demonstrate the application of this system for the high-level expression of plant, mammalian and bacterial proteins in E. coli.
外源基因在大肠杆菌中的表达需要原核转录和翻译元件与外源基因的编码区并列。通常情况下,这只会导致外源基因产物的适度表达。在此,我们描述了一种新型核糖体结合位点(RBS;噬菌体T7“基因10前导序列”),它能够驱动多个外源基因的翻译。这种RBS显著提高了我们迄今测试的所有基因的翻译效率,对外源基因尤其有效。增强后的表达通常比使用“通用”RBS时获得的表达高出40多倍。已构建了一种通用质粒载体,其中包含T7基因10前导序列,可方便地表达重要的基因产物。在本报告中,我们展示了该系统在大肠杆菌中高水平表达植物、哺乳动物和细菌蛋白质的应用。