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Besides the previously deduced 2X22-bp GInR binding boxes (gTnAc-n6-GaAAc-n6-GtnAC-n6-GAAAc-n6, briefly, a1-b1 and a2-b2), another 22-bp GInR binding box consisted of a3-site-n6-b3-site (briefly, a3-b3) was identified by DNase I footprinting assay in the promoter of Streptomyces coelicolor amtB, which overlapped with the PhoP binding sequences [1] Electrophoretic mobility shift assay (EMSA) employing purified recombinant GInR and the synthetic amtB promoter fragments with the three GInR binding boxes individually mutated,demonstrated that every box was involved in GInR binding in vitro.