Yamaichi Yoshiharu, Dörr Tobias
Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, CEA, CNRS, Université Paris Sud, 1 Avenue de la Terrasse, 91198, Gif sur Yvette, France.
Department of Microbiology, Weill Institute for Cell and Molecular Biology, Cornell University, Ithaca, NY, 14853, USA.
Methods Mol Biol. 2017;1624:39-49. doi: 10.1007/978-1-4939-7098-8_4.
Transposon insertion site sequencing (TIS) permits genome-wide, quantitative fitness assessment of individual genomic loci. In addition to the identification of essential genes in given growth conditions, TIS enables the elucidation of genetic networks such as synthetic lethal or suppressor gene combinations. Therefore, TIS becomes an exceptionally powerful tool for the high-throughput determination of genotype-phenotype relationships in bacteria. Here, we describe a protocol for the generation of high-density transposon insertion libraries and subsequent preparation of DNA samples for Illumina sequencing using the Gram-negative bacterium Vibrio cholerae as an example.
转座子插入位点测序（TIS）可对单个基因组位点进行全基因组范围的定量适应性评估。除了在特定生长条件下鉴定必需基因外，TIS还能阐明诸如合成致死或抑制基因组合等遗传网络。因此，TIS成为高通量确定细菌基因型-表型关系的一项极其强大的工具。在此，我们以革兰氏阴性菌霍乱弧菌为例，描述一种生成高密度转座子插入文库以及随后制备用于Illumina测序的DNA样本的方案。
