title: ARF-TSS: an alternative method for identification of transcription start site in bacteria.
abstract: Current methods for identifying transcription start sites (TSSs) of specific genes in bacteria usually require adaptors or radioactive labeling. These approaches can be technically demanding and environmentally unfriendly. Here we present a method for identifying TSS called ARF-TSS, which is based on cDNA generation, circularization, PCR amplification, and DNA sequencing to determine the 5'-end of transcripts, thus circumventing the need for adaptors and radioactive labeling. We validated the method using the gene lasI from the bacterial pathogen Pseudomonas aeruginosa. Our results show that ARF-TSS could be a good alternative to traditional methods for bacterial TSS analysis.
authors: Institute of Molecular and Cell Biology, Singapore.
authors: Chao
authors: C
authors: Wang
authors: Institute of Molecular and Cell Biology, Singapore.
authors: Jasmine
authors: J
authors: Lee
authors: Institute of Molecular and Cell Biology, Singapore.
authors: Yinyue
authors: Y
authors: Deng
authors: Institute of Molecular and Cell Biology, Singapore.
authors: Fei
authors: F
authors: Tao
authors: Institute of Molecular and Cell Biology, Singapore.
authors: Lian-Hui
authors: LH
authors: Zhang
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011550:Pseudomonas aeruginosa / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D024363:Transcription Initiation Site
chemical_list: D004269:DNA, Bacterial
keywords: Pseudomonas aeruginosa; mRNA; promoter; reverse transcription; transcriptional initiation; transcripts