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Upstream CRP-binding site is not essential for CRP-cAMP-mediated inhibition on the nifU promoter

查看全文 作  者:LI [1]Zhiting;ZHANG [1]Weijia;WANG [1]Yiping 高影响力作者 机构地区:[1]National Laboratory of Protein Engineering and Plant Genetic Engineering,College of Life Sciences,Peking University,Beijing 100871,China高影响力机构 出  处:《Chinese Science Bulletin》索引2002年第47卷第21期,共5页高影响力期刊 基  金:This work was supported in part by the National Natural Science Foundation of China (Grant No. 39925017), the International Cooperation Program (Grant No. 2001AA214211), the '863' Program (Grant No. 2001AA214021) and the '973' Program (Grant No. 2001C 摘  要:When the NifA-mediated activation of Kleb-siella pneumontae nifU promoter is recreated in Escherichia coli, it has been observed that CRP-cAMP has an inhibitory effect on the nifU promoter. Sequence analysis indicates that there is a strong CRP-binding site located upstream of the nifU promoter, overlapping completely with a previously identified Nif A-binding site. In vitro gel retardation analysis indicates that this putative CRP-binding site has similar affinity for CRP, when compared with that at the lac promoter, suggesting that CRP could effectively compete with NifA for such a binding site under physiological conditions. When this putative CRP-binding site on nifU was mutated, in vitro gel retardation analysis indicates that CRP can no longer bind to the mutant promoter. However, when constitutively expressed NifA is used as the activator, CRP-cAMP-mediated inhibitory effect on this mutant nifU promoter has no significant difference when compared with that obtained from its wild-type promoter. 关 键 词:CRP-cAMP the nifU PROMOTER gene regulation.
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