Reporter-Guided Transposon Mutant Selection for Activation of Silent Gene Clusters in Burkholderia thailandensis
Reporter-Guided Transposon Mutant Selection for Activation of Silent Gene Clusters in Burkholderia thailandensis
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DOI：
10.1002/cbic.201900748
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发表时间：
2020-03-03
期刊：
影响因子：
3.2
通讯作者：
Seyedsayamdost, Mohammad R.
中科院分区：
文献类型：
--
作者：
Mao, Dainan;Yoshimura, Aya;Seyedsayamdost, Mohammad R.
大多数天然产物生物合成基因簇，可以观察到的生物信息学是沉默的。这一见解促使了几种诱导其表达的方法的发展。最近的一种方法，称为突变体引导的突变体选择（RGMS），需要创建突变体库，然后通过报告基因表达筛选所需的表型。在此，我们采用类似的方法，以伯克霍尔德氏菌泰国，并使用转座子诱变，诱变三个菌株，每个携带荧光报告的malleilactone（mal），capistruin（cap），或一个身份不明的核糖体肽（tomm）基因簇。我们表明，即使是一个小的图书馆的马尔集群。转座子插入pyrF以及小分子介导的PyrF抑制都触发了鼻疽内酯的生物合成。我们的研究结果为RGMS和相关方法在伯克霍尔德氏菌中的广泛应用铺平了道路。
Most natural product biosynthetic gene clusters that can be observed bioinformatically are silent. This insight has prompted the development of several methodologies for inducing their expression. One of the more recent methods, termed reporter-guided mutant selection (RGMS), entails creation of a library of mutants that is then screened for the desired phenotype via reporter gene expression. Herein, we apply a similar approach to Burkholderia thailandensis and, using transposon mutagenesis, mutagenize three strains, each carrying a fluorescent reporter in the malleilactone (mal), capistruin (cap), or an unidentified ribosomal peptide (tomm) gene cluster. We show that even a small library of mal cluster. Both a transposon insertion into pyrF as well as small-molecule-mediated inhibition of PyrF trigger malleilactone biosynthesis. Our results pave the way toward the broad application of RGMS and related approaches to Burkholderia spp.