[A novel strategy for systematic identification of natural antisense transcripts of Pseudomonas aeruginosa based on RNase I protection assay].
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 systematic identified in high throughput for a lack of effective experimental method. Here, we have developed a strategy based on RNase I protection assay, which permit us to identify NATs in large scale which 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 random ly 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 be exist some gene regulatory mechanism acted by NATs in PAO1 and this worth further analysis.
Zhaohui
Yang
Y
Xiaolin
Jin
J
Xiancai
Rao
R
Xiaoxing
Cheng
C
Fuquan
Hu
H
D002874:Chromosome Mapping / Q000379:methods*; D019295:Computational Biology; D015723:Gene Library; D005821:Genetic Techniques; D023282:Nuclease Protection Assays / Q000379:methods*; D011550:Pseudomonas aeruginosa / Q000235:genetics*; D016372:RNA, Antisense / Q000737:chemistry / Q000302:isolation & purification*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D017423:Sequence Analysis, RNA; D014158:Transcription, Genetic
D016372:RNA, Antisense; D012259:Ribonuclease, Pancreatic