Heatwole V M, Somerville R L
Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907-6799.
J Bacteriol. 1992 Jan;174(1):331-5. doi: 10.1128/jb.174.1.331-335.1992.
Computer analysis identified a potential Trp repressor operator 56 nucleotides downstream of the transcriptional start point of aroL, the gene that encodes shikimate kinase II. Tryptophan-dependent interaction of Trp repressor with this operator was demonstrated in vitro by means of a restriction endonuclease protection assay. Regulation of expression from the aroL promoter was evaluated with several genetically marked Escherichia coli strains by using a single-copy aroL-lacZ transcriptional-translational reporter system. The expression of aroL was repressed 6.9-fold by the Tyr repressor alone and 29-fold when both Tyr and Trp repressors were present. The Trp repressor had no effect on expression from the aroL promoter in the absence of the Tyr repressor. Possible mechanisms for Trp repressor-mediated repression, including cooperative interactions with the Tyr repressor, are discussed.
计算机分析在编码莽草酸激酶II的基因aroL转录起始点下游56个核苷酸处鉴定出一个潜在的色氨酸阻遏物操纵子。通过限制性内切酶保护试验在体外证明了色氨酸阻遏物与该操纵子的色氨酸依赖性相互作用。利用单拷贝aroL-lacZ转录-翻译报告系统,用几种基因标记的大肠杆菌菌株评估了aroL启动子的表达调控。单独的酪氨酸阻遏物可使aroL的表达受到6.9倍的抑制,而当酪氨酸阻遏物和色氨酸阻遏物同时存在时,抑制倍数为29倍。在没有酪氨酸阻遏物的情况下,色氨酸阻遏物对aroL启动子的表达没有影响。本文讨论了色氨酸阻遏物介导的抑制作用的可能机制,包括与酪氨酸阻遏物的协同相互作用。