Olins P O, Rangwala S H
Department of Biological Sciences, Monsanto Company, St. Louis, Missouri 63198.
J Biol Chem. 1989 Oct 15;264(29):16973-6.
We recently reported that a ribosome binding site (RBS) derived from gene 10 of bacteriophage T7 (g10-L) causes a pronounced stimulation of expression when placed upstream of a variety of genes, and that this effect is probably due to a stimulation of translation efficiency in Escherichia coli (Olins, P. O., Devine, C. S., Rangwala, S. H., Kavka, K. S. (1988) Gene (Amst.) 73, 227-235). Here we present a model for the mechanism of action of the g10-L: the RBS contains a 9-base sequence which has the potential for forming a novel base-paired interaction with bases 458-466 of the 16 S rRNA of E. coli. Although such sequence homologies are rare in E. coli RBS regions, a number of similar sequences were found in the RBS regions of other bacteriophage structural genes. When an isolated homology sequence was placed upstream of a synthetic RBS, there was a 110-fold increase in the translation efficiency of the lacZ gene. Surprisingly, the homology sequence also stimulated translation when placed downstream of the initiator codon, indicating that this sequence is acting as a translational "enhancer."
我们最近报道,源自噬菌体T7基因10的核糖体结合位点(RBS,即g10-L),当置于多种基因上游时可显著刺激基因表达,且这种效应可能是由于其对大肠杆菌翻译效率的刺激作用(奥林斯,P.O.,迪瓦恩,C.S.,兰格瓦拉,S.H.,卡夫卡,K.S.(1988年)《基因》(阿姆斯特丹)73卷,227 - 235页)。在此我们提出一个关于g10-L作用机制的模型:该RBS包含一个9碱基序列,它有可能与大肠杆菌16 S rRNA的458 - 466位碱基形成一种新的碱基配对相互作用。尽管这种序列同源性在大肠杆菌RBS区域中很少见,但在其他噬菌体结构基因的RBS区域发现了许多类似序列。当一个分离的同源序列置于合成RBS上游时,lacZ基因的翻译效率提高了110倍。令人惊讶的是,该同源序列置于起始密码子下游时也能刺激翻译,这表明该序列起到了翻译“增强子”的作用。