Stanssens P, Remaut E, Fiers W
Cell. 1986 Mar 14;44(5):711-8. doi: 10.1016/0092-8674(86)90837-8.
Expression plasmids containing the E. coli lacZ coding region preceded by a set of different ribosome-binding sites and put under transcriptional control of the leftward promoter of phage lambda (PL) were used to study the synthesis of lacZ mRNA. In a normal host the steady state level of full-length lacZ mRNA varied 100-fold with the different synthesis levels of beta-galactosidase, whereas in a host expressing the antitermination protein N of phage lambda, all vectors synthesized the same amount of full-length lacZ mRNA, while maintaining the differences in beta-galactosidase expression. We present evidence for a causal relationship between the rate of ribosome loading and the continuation of transcription across the lacZ gene. We suggest that extended spacing between the RNA polymerase and the elongating ribosome causes transcriptional polarity by increasing the extent of premature termination. The conditional character of the termination event can best be explained by invoking termination factor Rho.
含有大肠杆菌lacZ编码区的表达质粒,其前面有一组不同的核糖体结合位点,并置于噬菌体λ左向启动子(PL)的转录控制之下,用于研究lacZ mRNA的合成。在正常宿主中,全长lacZ mRNA的稳态水平随β-半乳糖苷酶合成水平的不同而变化100倍,而在表达噬菌体λ抗终止蛋白N的宿主中,所有载体合成相同量的全长lacZ mRNA,同时保持β-半乳糖苷酶表达的差异。我们提供了核糖体装载速率与lacZ基因转录延续之间因果关系的证据。我们认为,RNA聚合酶与延伸核糖体之间的间隔延长会通过增加过早终止的程度而导致转录极性。终止事件的条件性特征最好通过引入终止因子Rho来解释。