Genome-wide transcription start site profiling in biofilm-grown Burkholderia cenocepacia J2315

Burkholderia cenocepacia is an opportunistic pathogenic bacterium intrinsically resistant to most antibiotics and biocides. The aim of this study is to map transcription start sites and 5'UTRs, and to discover novel non-coding small RNAs expressed in biofilms. The experimental approach used for this study is differential sequencing, where RNA samples are split into two aliquots, one of which is then treated with a 5' monophosphate-dependent exonuclease. Two separate libraries are created from exonuclease-treated and -untreated sub-samples, using illumina sequencing from the 5'end, without fractionation step and without depletion of abundant rRNAs. Transcription start sites can then be identified by comparing exonuclease-treated with untreated RNA-seq libraries.

洋葱伯克霍尔德菌（Burkholderia cenocepacia）是一类机会致病性细菌，对绝大多数抗生素及生物杀灭剂均具有固有耐药性。本研究旨在定位转录起始位点（transcription start sites）与5'非翻译区（5'UTRs），并发掘生物被膜中表达的新型非编码小RNA。本研究采用差异测序策略：将RNA样品均分为两份，其中一份经5'单磷酸依赖性核酸外切酶处理；分别以经核酸外切酶处理与未处理的子样品构建测序文库，采用Illumina测序技术对5'端进行测序，无需分级分离步骤，亦未对丰度较高的核糖体RNA进行去除。通过对比经核酸外切酶处理与未处理的RNA测序文库，即可鉴定转录起始位点。