Goliger J A, Yang X J, Guo H C, Roberts J W
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, NY 14853.
J Mol Biol. 1989 Jan 20;205(2):331-41. doi: 10.1016/0022-2836(89)90344-6.
We have constructed novel transcription templates in which we have fused the late gene promoters of Escherichia coli phages lambda and 82 upstream from three different rho-independent transcription terminators. Using an in vitro transcription assay and an in vivo galactokinase expression assay, we find that the initial portion of the transcribed region significantly affects the efficiency of some downstream terminators. We have identified, by deletion, substitution and point mutation analysis, sequences responsible for these increased levels of factor-independent readthrough. Since these important sequences occur within about 30 nucleotides of the RNA start site, we suggest that the initial portion of the transcript can affect termination efficiency.
我们构建了新型转录模板,其中将大肠杆菌噬菌体λ和82的晚期基因启动子融合在三个不同的不依赖ρ因子的转录终止子上游。通过体外转录测定和体内半乳糖激酶表达测定,我们发现转录区域的起始部分显著影响一些下游终止子的效率。我们通过缺失、替换和点突变分析确定了导致这些不依赖因子通读水平增加的序列。由于这些重要序列出现在RNA起始位点约30个核苷酸范围内,我们认为转录本的起始部分会影响终止效率。