Department of Biology and Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON, Canada.
Methods Mol Biol. 2022;2489:157-171. doi: 10.1007/978-1-0716-2273-5_9.
Bacteria produce an impressive array of bioactive specialized metabolites, with Streptomyces (and the actinobacteria more generally) being unusually diverse and prolific producers. However, the biosynthetic potential of these organisms has yet to be fully explored, as many of the biosynthetic gene clusters that direct the synthesis of these natural products are transcriptionally silent under laboratory growth conditions. Here, we describe strategies that can be employed to broadly stimulate the expression of biosynthetic gene clusters in Streptomyces and their relatives, follow the transcription of these genes, and assess the antimicrobial activity of the resulting molecules.
细菌产生了令人印象深刻的一系列生物活性特殊代谢产物,链霉菌(和更广泛的放线菌)是异常多样化和丰富的生产者。然而,这些生物体的生物合成潜力尚未得到充分探索,因为许多指导这些天然产物合成的生物合成基因簇在实验室生长条件下转录沉默。在这里,我们描述了可以广泛用于刺激链霉菌及其亲缘种生物合成基因簇表达的策略,跟踪这些基因的转录,并评估产生的分子的抗菌活性。