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Mentz, Almut; Neshat, Armin; Pfeifer-Sancar, Katharina; Pühler, Alfred ; Rückert, Christian ; Kalinowski, Jörn: Comprehensive discovery and characterization of small RNAs in Corynebacterium glutamicum ATCC 13032. In: BMC Genomics. Jg.14 H. 1. 2013
Inhalt
Results
Detection of potential sRNA genes in C. glutamicum ATCC 13032 by transcriptome sequencing and read-mapping
Characterization of potential sRNA genes with the help of bioinformatic promoter analysis
Classification of potential sRNA genes by their positions relative to annotated protein-coding sequences
Detection of small C. glutamicum genes encoding small proteins
Analysis of cis-antisense RNA genes, located within protein-coding genes
Bioinformatic analysis of sequence and structural conservation of trans-encoded sRNAs in C. glutamicum ATCC 13032
Experimental validation of three trans-encoded sRNAs by Northern blotting of stress-specific RNA samples and detailed structure analysis
Discussion
Conclusions
Methods
Preparation of cDNA libraries for RNA-Sequencing
Bacterial growth conditions and total RNA-isolation
Preparation of two different cDNA libraries for sequencing
Bioinformatics analysis
Read mapping and data visualization
Detection of transcription start sites
Promoter search using primary 5′-end data
Rho-independent terminator search
ORF and RBS prediction
Prediction of secondary structure conservation
Other tools and software
Northern blot
Additional files
Abbreviations
Competing interests
Authors’ contributions
Acknowledgements
Author details
References