The time course analysis of TNM A production in Streptomyces sp. CB03234 wild-type and CB03234-R-16 mutant, and qRT-PCR analysis revealed the overexpression of key genes in Streptomyces sp. CB03234-R-16, comparing to the wild-type. a The biosynthetic gene cluster of TNM A in Streptomyces sp. CB03234 and the genes [5] evaluated in qRT-PCR were highlighted in red. b The production time course of TNM A in Streptomyces sp. CB03234 and Streptomyces sp. CB03234-R-16 in shaking flasks. c–j qRT-PCR analysis of relA and selected genes in TNM A biosynthetic gene cluster. The white bar stands for Streptomyces sp. CB03234 wild-type, and the red bar stands for CB03234-R-16 mutant. hrdB was used as an internal control. tnmL, cytochrome P450 monooxygenase; tnmR7, AraC family transcriptional regulator; tnmR3, unknown protein; tnmE10, type II thioesterase; tnmR1, HxlR family transcriptional regulator; tnmR2, putative regulator; tnmR4, AraC family transcriptional regulator. Each data point represents the mean ± SD of three biological replicates (color figure online)