The products of most secondary metabolite biosynthetic gene clusters (BGCs) have yet to be discovered, in part due to low expression levels in laboratory cultures. Reporter-guided mutant selection (RGMS) has recently been developed for this purpose: a mutant library is generated and screened, using genetic reporters to a chosen BGC, to select transcriptionally active mutants that then enable the characterization of the “cryptic” metabolite. The requirement for genetic reporters limits the approach to a single pathway within genetically tractable microorganisms. Herein, we utilize untargeted metabolomics in conjunction with transposon mutagenesis to provide a global read-out of secondary metabolism across large numbers of mutants. We employ self-organizing map analytics and imaging mass spectrometry to identify and characterize seven cryptic metabolites from mutant libraries of two different Burkholderia species. Applications of the methodologies reported can expand our understanding of the products and regulation of cryptic BGCs across phylogenetically diverse bacteria.

中文翻译：

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使用质谱引导的转座子突变体选择解锁隐秘代谢物。

大多数次生代谢物生物合成基因簇 (BGC) 的产物尚未被发现，部分原因是实验室培养物中的表达水平较低。最近为此目的开发了报告基因引导的突变体选择 (RGMS)：生成和筛选突变体文库，使用选定的 BGC 的遗传报告基因，选择具有转录活性的突变体，然后对“神秘”代谢物进行表征。对遗传记者的要求将方法限制在遗传易处理微生物中的单一途径。在此，我们利用非靶向代谢组学与转座子诱变相结合，提供大量突变体的次级代谢的全局读数。伯克霍尔德菌种。报告的方法的应用可以扩大我们对系统发育多样化细菌中神秘 BGC 的产品和调控的理解。