A genetic screen suggests an alternative mechanism for inhibition of SecA by azide

A high-density transposon library constructed in BW25113 was grown in the absence or presence of 0.25 mM or 0.5 mM sodium azide. In the case of the 0 mM and 0.25 mM samples, cells were grown until OD600 = 1.0. In the case of of the 0.5 mM sodium azide sample, the cells were grown to OD600 = 0.9, at which point the cells stopped growing exponentially. The location of the transposons after growth was determined using Illumina sequecing. Libraries were prepared using arbirary PCR and sequenced using single-end reads. Reads were then processed an aligned to the genome sequence for Escherichia coli K-12 strain W3110 (NCBI accession AP009048.1). Please see associated manuscript at BiorXiv for a more detailed description of the experiment.本研究在BW25113菌株中构建了高密度转座子文库（transposon library），并设置无添加、0.25 mM及0.5 mM叠氮化钠三种培养条件。对于0 mM与0.25 mM组样品，菌体培养至OD600值为1.0时终止培养；0.5 mM叠氮化钠组样品则培养至OD600值为0.9时，菌体即停止指数生长。培养完成后，采用Illumina测序（Illumina sequencing）技术对转座子的插入位置进行鉴定。文库构建采用任意PCR（arbitrary PCR）方法，测序采用单端读长（single-end reads）策略。随后对测序读段进行处理并比对至大肠杆菌K-12菌株W3110的基因组序列（NCBI登录号：AP009048.1）。有关本实验的详细描述，请参见预印本平台BioRxiv上的相关稿件。