GlnR has been characterized as a central regulator governing most nitrogen metabolisms in many important actinomycetes. So far, the GlnR binding consensus sequences have been extensively studied, but with different motifs proposed, which has therefore brought confusion and impeded the understanding of the in-depth molecular mechanisms of GlnR-mediated transcriptional regulation. Here, a 30-nt GlnR-protected DNA sequence in the promoter of ginA in Amycolatopsis mediterranei was employed for precise characterization of GlnR binding consensus sequences. Site-by-site mutagenesis strategy combining with the Electrophoretic Mobility Shift Assay were employed, and a 5-nt GlnR Box was precisely defined as the basic unit for GInR binding. (C) 2015 Elsevier Inc. All rights reserved.
[1]Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, CAS Key Lab Synthet Biol, Shanghai, Peoples R China;
[2]Fudan Univ, Sch Life Sci, Dept Microbiol & Microbial Engn, State Key Lab Genet Engn, Shanghai 200433, Peoples R China;
[3]Chinese Natl Human Genome Ctr, Shanghai MOST Key Lab Hlth & Dis Genom, Shanghai, Peoples R China;
[4]Chinese Univ Hong Kong, Prince Wales Hosp, Dept Microbiol, Shatin, Hong Kong, Peoples R China;
[5]Chinese Univ Hong Kong, Prince Wales Hosp, Li Ka Shing Inst Hlth Sci, Shatin, Hong Kong, Peoples R China
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