A Novel Strategy for Systematic Identification of Natural Antisense Transcripts of Pseudomonas aeruginosa Based on the RNase I Protection Assay

摘要

Natural antisense transcripts (NATs) are widespread in prokaryotes and eukaryotes. They have very important functions in regulating expression of their target genes. However, until now, NATs haven't been systematically identified in high throughput for a lack of an effective experimental method. Here, we have developed a strategy based on the RNase I protection assay, which permits us to identify NATs in large scale which is presented inside Pseudomonas aeruginosa. After being isolated from total RNA of P. aeruginosa strain ATCC 6872 (PAO1), NATs that encoded chromosomally were cloned in a simple and efficient way and a NATs gene library was constructed successfully. 78 randomly selected positive clones were sequenced and analyzed by bioinformatic tools. There are several candidate molecules which are located in PAO1 genome precisely. This study suggests that there may exist some gene regulatory mechanism acted by NATs in PAO1 and this is worth further analysis.