RNA-seq in polysome fractions

We previously reported that ribosome-targeting antibiotic lincomcyin at concentrations below the minimum inhibitory concentrations (MIC) enhance the secondary metabolism in streptomycetes. Through the work, we have revealed that qualitative and quantitative changes in ribosomes may be a key to lincomcyin-induced secondary metabolism in Streptomyces coelicolor A3(2). In this project, gene expression analysis of mRNA in polysomal fractions was performed to analyze the effect of altered ribosomes in the presence of lincomycin on their translational properties. RNA was prepared from polysomal fractions in the absence and presence of lincomycin and subjected to RNA-seq. Since polysomes contain mRNAs that are exactly in translation, it is possible to determine which genes are actively translated in the presence of lincomycin.