Wright J J, Hayward R S
EMBO J. 1987 Apr;6(4):1115-9. doi: 10.1002/j.1460-2075.1987.tb04866.x.
We have examined the possibility that translation reading through a fully rho-independent transcriptional terminator in Escherichia coli might prevent termination, as already established for rho-dependent terminators. Plasmids were constructed with and without interposition of the rho-independent coliphage T7 'early' terminator between a promoter and galK. Our constructions ensured either that there was no upstream translation, or that translation (initiated at the galE ribosome binding site) stopped upstream of, or at the normal position (the T7 gene 1.3 stop codon) with respect to, the transcriptional terminator; or else downstream of both this stop codon and the terminator. Our galactokinase enzyme and mRNA measurements on strains harbouring these plasmids indicate that 'readthrough translation' eliminates transcriptional termination at the T7 site. This effect is suppressed if the rate of ribosome movement is reduced with fusidic acid.
我们已经研究了在大肠杆菌中通过完全不依赖ρ因子的转录终止子进行翻译通读可能阻止终止的可能性,就像已确定的依赖ρ因子的终止子那样。构建了一些质粒,在启动子和galK之间分别插入或不插入不依赖ρ因子的噬菌体T7“早期”终止子。我们的构建确保要么不存在上游翻译,要么翻译(在galE核糖体结合位点起始)在转录终止子的上游、正常位置(T7基因1.3的终止密码子)处或终止密码子和终止子两者的下游停止。我们对携带这些质粒的菌株进行的半乳糖激酶酶活性和mRNA测量表明,“通读翻译”消除了T7位点的转录终止。如果用夫西地酸降低核糖体移动速率,这种效应就会受到抑制。