Tobacco Smoke Exacerbates Filifactor alocis Pathogenicity. Tobacco Smoke Exacerbates Filifactor alocis Pathogenicity

RNA sequencing was used to identify genes up- and down-regulated when Fa was grown in cigarette smoke media Overall design: F alocis grown in BHI in anaerobic conditions was passaged into BHI or CSE (BHI with smoke extract to a concentration of 1000ng/ml nicotine) at mid log stage. RNA from original culture and from each second passage condition was prepared using MP Fast Prep and RNeasy kit. RNA seq was performed on each passage and up and down-regulated genes unique in passage from BHI to CSE were analysed.

本研究采用RNA测序技术，鉴定Fa在香烟烟雾培养基中培养时的上调与下调基因。
实验设计：将在厌氧条件下于脑心浸液培养基（Brain Heart Infusion, BHI）中培养的F alocis在对数生长期中期转接至BHI培养基或香烟烟雾提取物（Cigarette Smoke Extract, CSE）培养基（即添加烟雾提取物且尼古丁终浓度为1000ng/ml的BHI培养基）中。使用MP Fast Prep样品制备系统与RNeasy试剂盒，分别制备原始培养物与各第二代传代培养条件下样本的总RNA。对各传代样本进行RNA测序，并分析从BHI培养基转接至CSE培养基的样本中特有的上调及下调基因。