High-throughput insertion tracking by deep sequencing for the analysis of bacterial pathogens.
High-throughput insertion tracking by deep sequencing for the analysis of bacterial pathogens.
复制标题
DOI：
10.1007/978-1-61779-089-8_15
复制
发表时间：
2011
影响因子：
--
通讯作者：
Sandy M. S. Wong;Jeffrey D. Gawronski;David S. Lapointe;B. Akerley
中科院分区：
文献类型：
--
作者：
Sandy M. S. Wong;Jeffrey D. Gawronski;David S. Lapointe;B. Akerley
用于鉴定感染期间生存和生长所需的微生物基因的全基因组技术对于细菌发病机制的研究非常有用。大规模并行测序平台的出现创造了显着加速此类基因组规模分析并实现前所未有的灵敏度、分辨率和定量的机会。本章概述了基因组规模的方法，该方法将高密度转座子诱变与 amariner 转座子和深度测序相结合，以识别发病机制实验模型中生存所需的基因。将这种方法应用于模型病原体流感嗜血杆菌，可以全面分析这种人类呼吸道病原体的每个基因在小鼠肺部模型中的相对作用。该方法很容易适用于几乎任何适合转座子诱变的生物体。
Whole-genome techniques toward identification of microbial genes required for their survival and growth during infection have been useful for studies of bacterial pathogenesis. The advent of massively parallel sequencing platforms has created the opportunity to markedly accelerate such genome-scale analyses and achieve unprecedented sensitivity, resolution, and quantification. This chapter provides an overview of a genome-scale methodology that combines high-density transposon mutagenesis with amarinertransposon and deep sequencing to identify genes that are needed for survival in experimental models of pathogenesis. Application of this approach to a model pathogen,Haemophilus influenzae, has provided a comprehensive analysis of the relative role of each gene of this human respiratory pathogen in a murine pulmonary model. The method is readily adaptable to nearly any organism amenable to transposon mutagenesis.