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Background  Identifying the regulatory factors that control transcriptional activity is a major challenge of gene expression studies. Here, we describe the application of a novel approach for in vivo identification of regulatory proteins that may directly or indirectly control the transcription of a promoter of interest in  Streptomyces .    Results  A method based on the combination of Tn5 minitransposon-driven random mutagenesis and  lux  reporter genes was applied for the first time for the  Streptomyces  genus. As a proof of concept, we studied the  topA  supercoiling-sensitive promoter, whose activity is dependent on unknown regulatory factors. We found that the  sco4804  gene product positively influences  topA  transcription in  S. coelicolor , demonstrating SCO4804 as a novel player in the control of chromosome topology in these bacteria.    Conclusions  Our approach allows the identification of novel  Streptomyces  regulators that may be critical for the regulation of gene expression in these antibiotic-producing bacteria.    Supplementary Information  The online version contains supplementary material available at 10.1186/s12934-021-01590-7.

Combining transposon mutagenesis and reporter genes to identify novel regulators of the topA promoter in Streptomyces