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首页> 外文期刊>PLOS Neglected Tropical Diseases >Genome-scale analysis of the genes that contribute to Burkholderia pseudomallei biofilm formation identifies a crucial exopolysaccharide biosynthesis gene cluster
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Genome-scale analysis of the genes that contribute to Burkholderia pseudomallei biofilm formation identifies a crucial exopolysaccharide biosynthesis gene cluster

机译：对Burkholderia pseudomallei生物膜形成有助于贡献的基因的基因组规范分析鉴定了一个关键的外偶糖生物合成基因簇

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Burkholderia pseudomallei, the causative agent of melioidosis, is an important public health threat due to limited therapeutic options for treatment. Efforts to improve therapeutics for B. pseudomallei infections are dependent on the need to understand the role of B. pseudomallei biofilm formation and its contribution to antibiotic tolerance and persistence as these are bacterial traits that prevent effective therapy. In order to reveal the genes that regulate and/or contribute to B. pseudomallei 1026b biofilm formation, we screened a sequence defined two-allele transposon library and identified 118 transposon insertion mutants that were deficient in biofilm formation. These mutants include transposon insertions in genes predicted to encode flagella, fimbriae, transcriptional regulators, polysaccharides, and hypothetical proteins. Polysaccharides are key constituents of biofilms and B. pseudomallei has the capacity to produce a diversity of polysaccharides, thus there is a critical need to link these biosynthetic genes with the polysaccharides they produce to better understand their biological role during infection. An allelic exchange deletion mutant of the entire B. pseudomallei biofilm-associated exopolysaccharide biosynthetic cluster was decreased in biofilm formation and produced a smooth colony morphology suggestive of the loss of exopolysaccharide production. Conversely, deletion of the previously defined capsule I polysaccharide biosynthesis gene cluster increased biofilm formation. Bioinformatics analyses combined with immunoblot analysis and glycosyl composition studies of the partially purified exopolysaccharide indicate that the biofilm-associated exopolysaccharide is neither cepacian nor the previously described acidic exopolysaccharide. The biofilm-associated exopolysaccharide described here is also specific to the B. pseudomallei complex of bacteria. Since this novel exopolysaccharide biosynthesis cluster is retained in B. mallei, it is predicted to have a role in colonization and infection of the host. These findings will facilitate further advances in understanding the pathogenesis of B. pseudomallei and improve diagnostics and therapeutic treatment strategies.
机译：Burkholderia Pseudomallei是杂种中的致病因子，是由于治疗有限的治疗选择，是一种重要的公共卫生威胁。改善B.Pseudomallei感染治疗方法的努力取决于了解B.Pseudomallei生物膜形成的作用以及其对抗生素耐受性和持久性的贡献，因为这些细菌性状可防止有效治疗。为了揭示调节和/或贡献B.Pseudomallei 1026b Biofilm的基因，我们筛选了一系列定义的双等位基因转座子文库，并鉴定了缺乏生物膜形成的118个转座子插入突变体。这些突变体包括预测鞭毛，FiMbriae，转录调节剂，多糖和假想蛋白的基因中的转座子插入。多糖是生物膜的关键成分，B.Pseudomallei具有产生多糖的多样性的能力，因此需要将这些生物合成基因与它们产生的多糖联系起来以更好地了解其在感染期间的生物学作用。生物膜形成下降了整个B.Pseudomallei生物膜相关的外偶多糖生物合成簇的等位基因交换缺失突变体。暗示了外偶糖类损失的光滑菌落形态。相反，缺失先前定义的胶囊I多糖生物合成基因簇的增加生物膜形成。生物信息学分析结合免疫斑分析和部分纯化的外偶糖的糖基组合物研究表明，生物膜相关的外偶糖既不是冰川也不是先前描述的酸性外偶。这里描述的生物膜相关的外偶糖也是特异于细菌的B.Pseudomallei复合物。由于这种新的外偶糖生物合成簇保留在B.Mallei中，因此预计在宿主的定植和感染中具有作用。这些发现将有助于了解B.Pseudomallei的发病机制，改善诊断和治疗策略的进一步进展。

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