Ion torrent-based transcriptional assessment of a Corynebacterium pseudotuberculosis equi strain reveals denaturing high-performance liquid chromatography a promising rRNA depletion method
| Autor: | Castro, Thiago L P, Seyffert, Nubia, Ramos, Rommel T J, Barbosa, Silvanira, Carvalho, Rodrigo D O, Pinto, Anne Cybelle, Carneiro, Adriana Ribeiro, Silva, Wanderson Marques, Pacheco, Luis G C, Downson, Christopher, Schneider, Maria P C, Miyoshi, Anderson, Azevedo, Vasco, Silva, Artur |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Corynebacterium Infections
Corynebacterium pseudotuberculosis High-Throughput Nucleotide Sequencing Nucleic Acid Denaturation QR RNA Bacterial QH301 Bacterial Proteins Species Specificity RNA Ribosomal Animals Horse Diseases Horses Transcriptome QH426 Research Articles Chromatography High Pressure Liquid |
| Zdroj: | Repositório Institucional da UFBA Universidade Federal da Bahia (UFBA) instacron:UFBA Microbial Biotechnology |
| ISSN: | 1751-7907 |
| DOI: | 10.1111/1751-7915.12020 |
| Popis: | Texto completo: acesso restrito. p. 168–177 Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2015-01-05T12:57:59Z No. of bitstreams: 1 Thiago L. P. Castro.pdf: 601454 bytes, checksum: e531120e79c4cdbf3d28a04a9292463e (MD5) Approved for entry into archive by Patricia Barroso (pbarroso@ufba.br) on 2015-05-05T14:22:40Z (GMT) No. of bitstreams: 1 Thiago L. P. Castro.pdf: 601454 bytes, checksum: e531120e79c4cdbf3d28a04a9292463e (MD5) Made available in DSpace on 2015-05-05T14:22:40Z (GMT). No. of bitstreams: 1 Thiago L. P. Castro.pdf: 601454 bytes, checksum: e531120e79c4cdbf3d28a04a9292463e (MD5) Previous issue date: 2013 Corynebacterium pseudotuberculosis equi is a Gram-positive pathogenic bacterium which affects a variety of hosts. Besides the great economic losses it causes to horse-breeders, this organism is also known to be an important infectious agent to cattle and buffaloes. As an outcome of the efforts in characterizing the molecular basis of its virulence, several complete genome sequences were made available in recent years, enabling the large-scale assessment of genes throughout distinct isolates. Meanwhile, the RNA-seq stood out as the technology of choice for comprehensive transcriptome studies, which may bring valuable information regarding active genomic regions, despite of the still impeditive associated costs. In an attempt to increase the use of generated reads per instrument run, by effectively eliminating unwanted rRNAs from total RNA samples without relying on any commercially available kits, we applied denaturing high-performance liquid chromatography (DHPLC) as an alternative method to assess the transcriptional profile of C. pseudotuberculosis. We have found that the DHPLC depletion method, allied to Ion Torrent sequencing, allows mapping of transcripts in a comprehensive way and identifying novel transcripts when a de novo approach is used. These data encourage us to use DHPLC in future transcriptional evaluations in C. pseudotuberculosis. |
| Databáze: | OpenAIRE |
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