Defining the ABC of gene essentiality in streptococci
Defining the ABC of gene essentiality in streptococci
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DOI：
10.1186/s12864-017-3794-3
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发表时间：
2017-05-31
期刊：
影响因子：
4.4
通讯作者：
Waller, Andrew S.
中科院分区：
文献类型：
--
作者：
Charbonneau, Amelia R. L.;Forman, Oliver P.;Waller, Andrew S.
背景：利用下一代测序来询问饱和细菌突变文库，为全基因组基因必要性的分配提供了前所未有的信息。将饱和突变文库暴露于特定条件和随后的测序可以用来揭示与该条件相关的基因本质。在这里，我们提出了一个条形码转座子定向插入位点测序（TraDIS）系统来定义马链球菌亚种的必要基因列表。马，马的勒死的病原体，第一次。将C组链球菌的基因必需性数据与A组和B组链球菌的基因必需性数据进行比较。神经突变文库包含33,000-66,000个独特的突变体。对从每个文库中提取的DNA进行TraDIS，并将数据单独分析或作为组合主库进行分析。基因的重要性决定了19.5%的equi基因组是必不可少的。与A组和B组链球菌的基因特征进行比较，3种链球菌的基因一致性分别为90.2%和89.4%，总体一致性为83.7%。结论：利用pGh9: ISS1条形码生成突变文库，为马链球菌和其他链球菌的基因功能定位提供了一个非常有用的工具。A、B和C群链球菌共有的基本基因集进一步证明了这些重要致病菌之间的密切遗传关系。因此，本文报道的基因重要性ABC为报道链球菌的功能基因组提供了坚实的基础。
Background: Utilising next generation sequencing to interrogate saturated bacterial mutant libraries provides unprecedented information for the assignment of genome-wide gene essentiality. Exposure of saturated mutant libraries to specific conditions and subsequent sequencing can be exploited to uncover gene essentiality relevant to the condition. Here we present a barcoded transposon directed insertion-site sequencing (TraDIS) system to define an essential gene list for Streptococcus equi subsp. equi, the causative agent of strangles in horses, for the first time. The gene essentiality data for this group C Streptococcus was compared to that of group A and B streptococci.nerate mutant libraries containing between 33,000-66,000 unique mutants. TraDIS was performed on DNA extracted from each library and data were analysed separately and as a combined master pool. Gene essentiality determined that 19.5% of the S. equi genome was essential. Gene essentialities were compared to those of group A and group B streptococci, identifying concordances of 90.2% and 89.4%, respectively and an overall concordance of 83.7% between the three species.Conclusions: The use of barcoded pGh9: ISS1 to generate mutant libraries provides a highly useful tool for the assignment of gene function in S. equi and other streptococci. The shared essential gene set of group A, B and C streptococci provides further evidence of the close genetic relationships between these important pathogenic bacteria. Therefore, the ABC of gene essentiality reported here provides a solid foundation towards reporting the functional genome of streptococci.