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. 2020 Jan 20;48(3):1583–1598. doi: 10.1093/nar/gkz1220

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Deletion of cdgB_gh_,rmdB_gh and bldD_gh strongly influences expression of genes essential for moenomycin production. (A) Genetic organization of moe cluster 1. (B) Comparison of ΔcdgB_gh and ATCC14672 transcriptional profiles. (C) Comparison of ΔrmdB_gh and ATCC14672 transcriptional profiles. (D) Comparison of ΔbldD_gh and ATCC14672 transcriptional profiles. The expression of tested genes was analyzed in 48 h cultures grown in TSB; 200 ng of RNA sample were used per reaction; C⁺, positive control (genomic DNA of ATCC14672 strain). Attempts to synthesize hrdB from RNA without pretreatment with RT served as negative controls (marked as C⁻). Total RNA samples were isolated from three independent biological replicates. The images represent the typical result of three independent RT-PCR experiments. (E) Transcriptional activity of the moeE5 promoter in Streptomyces ghanaensis strains. (F) Transcriptional activity of the adpA_gh promoter in S. ghanaensis strains. (G) Transcriptional activity of the bldA_gh promoter in S. ghanaensis strains. WT-C, ΔcdgB-C, ΔrmdB-C and ΔbldD-C: control strains carrying promoterless vector pGUS. Cultures and subsequent β-glucuronidase assays were performed in triplicate. Values were normalized to equal amounts of dry biomass. Error bars, ±2 SD. Significance of difference in the transcriptional activities of tested promoters was calculated by a two-tailed t-test. Asterisks indicate the significance value (^∗P < 0.05, ^∗∗P < 0.01).