Análise bioinformática do perfil de transcritos Klebsiella pneumoniae através de dados de rna-seq

• Autor(a) principal: Custódio, Márlon Grégori Flores
• Data de Publicação: 2015
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Biblioteca Digital de Teses e Dissertações do LNCC
• Texto Completo: https://tede.lncc.br/handle/tede/231
• topic: Bioinformática; Bioinformatics; CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL; CNPQ::CIENCIAS EXATAS E DA TERRA::CIENCIA DA COMPUTACAO::MATEMATICA DA COMPUTACAO::MODELOS ANALITICOS E DE SIMULACAO

The indiscriminate use of antibiotics or their incorrect administration has made over the years, these drugs are losing their effectiveness due to evolutionary mechanisms, which naturally confer resistance characteristics to bacterias. As example of this evolutionary mechanism, we can mention the recent reports polymyxins resistant bacteria, drugs used as a last resort in infection control by super resistant bacteria, such as bacteria of the species Klebsiella pneumoniae microarray. The ease of this bacterium in performing transfers of genetic material, together with other mechanisms of their own group, is also making it resistant to polymyxins. This bacterial resistance points to a serious epidemiological problem, causing deaths including in Brazil. This study, aimed to infer the possible metabolic pathways and genes active in gene regulation mechanism related to resistance to polymyxin B in the genome of the bacteria K. pneumoniae KP13 strain, which had its entire genome unraveled in 2013. This opportunistic pathogenic microorganism He was responsible for hospital infection outbreak in 2009 in the south of the country. The findings of this study were made from the RNA-Seq technique, which is a transcriptome analysis technique based on the next generation sequencing (NGS), which allows a review of gene expression on a large scale. The transcriptome study, among others gives us an overview of the set of messengers transcripts (mRNAs) in a cell, and this allows us to directly evaluate the expression of its genes in specific situations. The transcriptome of the study was examined body 6 under conditions with two biological replicates for each condition. For sequencing were used two sequencing platforms: Illumina HiSeq and Roche 454 which enabled a comparative analysis of the data. From the obtained result of gene expression, the first stage of the study was the pre-processing of RNA-Seq data, where it was developed a methodology that was the basis for this analysis prokaryotic organism, given that the vast majority of materials available aims to study eukaryotic organisms. In the second stage of labor, the alignments were generated and the following was made to quantify the expression for each gene of the bacteria under study. The next step was to examine differential expression of genes important step towards the elucidation of resistance targets of regulation. All the differential expression of genes procedure was done using the R platform and the EDGE R package, the most suitable for the size of data that would be analyzed. The inference of genes active in gene regulation mechanism related to resistance to polymyxin B in the genome of the bacteria K. pneumoniae KP13 was made based on the clustering technique k-means, which showed to be effective within the universe of data to be mined. For the data generated, we were obtained $ 150 $ groups from the set 70 % genes most differentially expressed in all study conditions and, of these, the most significant associated with drug resistance and their metabolic pathways were chosen were investigated. Groupings proved concise and technical shows stable for application tests with other bodies.