Random transposon mutagenesis of Verrucomicrobium spinosum DSM 4136T
Random transposon mutagenesis of Verrucomicrobium spinosum DSM 4136T
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DOI：
10.1007/s00203-010-0666-5
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发表时间：
2011-04-01
影响因子：
2.8
通讯作者：
Ward, Naomi L.
中科院分区：
文献类型：
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作者：
Domman, Daryl B.;Steven, Blaire T.;Ward, Naomi L.
由于它们作为自由生活和与宿主相关的微生物在环境中无处不在，所以Verrucommicrobia是一个越来越受关注的细菌群体。它们还展示了一种不同寻常的分区细胞计划，与包括Planctomycete细菌在内的邻近门的成员共享。然而，由于缺乏现有的遗传工具来对从生态和基因组数据中提出的假设进行强有力的测试，目前很难对Verrucommicrobia进行研究。据我们所知，还没有发表的研究描述外源DNA转化到Verrucommicrobia(或邻近的含有Planctomycens的门)的任何成员。在这里，我们提出了一种通过电穿孔将DNA转化到棘毛韦氏杆菌DSM 4136(T)的方法，并首次描述了这种生物中的一个随机转座子突变体文库。我们预计，这种方法可以成功地应用于其他Verrucommicrobia，提供机会来测试预测的基因功能在生态相互作用中的作用，并确定与独特的Planctomycete-Verrucom微生物细胞计划相关的基因。
The Verrucomicrobia are a bacterial group of growing interest due to their environmental ubiquity as free-living and host-associated microbes. They also exhibit an unusual compartmentalized cell plan, shared with members of neighboring phyla that include the Planctomycete bacteria. However, Verrucomicrobia are currently difficult to study, due to a lack of available genetic tools that would permit robust testing of hypotheses formulated from ecological and genomic data. To our knowledge, there are no published studies describing the transformation of exogenous DNA into any members of the Verrucomicrobia (or the neighboring phylum containing Planctomycetes). Here, we present a procedure for the transformation of DNA into Verrucomicrobium spinosum DSM 4136(T) via electroporation and the first description of a random transposon mutant library in this organism. We anticipate that this approach could be applied successfully to other Verrucomicrobia, providing opportunities to test the role of predicted gene function in ecological interactions and identify genes associated with the distinctive Planctomycete-Verrucomicrobial cell plan.