Project description:The immune response associated with mastitis caused by Mycoplasma bovis is a very complicated biological process in several type of cells, including immune cells, mammary epithelial cells and, endothelial cells. Thus, revealing of the microRNAs in the Mycoplasma bovis infected mammary gland tissues is particularly important for the immune response mechanism to Mycoplasma bovis. Firstly, mammary gland tissue samples were collected from Holstein cows and screened for Mycoplasma bovis. Then, total RNA was isolated from mycoplasma bovis infected tissues and RNA sequencing was performed. After bioinformatics analysis, GO and KEGG analysis of target genes of identified microRNAs were conducted. Our results revaled that 24 of the known microRNAs were expressed differently and 13 of the novel microRNAs were expressed differently in Mycoplasma bovis positive tissues. The target genes of these microRNAs were found to be associated with especially inflammation pathways. In conclusion, this study demonstrated that identified miRNAs may be involved in the signaling pathways during mastitis case caused by Mycoplasma bovis.
Project description:Mycoplasma gallisepticum transcriptome comparison between in vitro grown cultures of strains Rlow and F utilizing oligo DNA microarrays.
Project description:Analysis of H292 cells infected with Mycoplasma hyorhinis. Mycoplasma infection reduces the cytotoxic effect of Nutlin3 on H292 cells. The results provide insight into molecular mechanisms underlying the response of H292 cells to Nutlin3.
Project description:Various anti-mycoplasma drugs have different effects on cells growth We used microarrays to detail the global programme of gene expression underlying gastric cancer cells treated with anti-mycoplasma drugs and identified distinct classes of up-regulated genes during this process.
Project description:The goal of this experiment is to determine the overall relative strength of promoter sequences in Mycoplasma feriruminatoris. For this, 2 replicates were grown in parallel in Hayflick media and the RNA wa extracted at exponential growth phase (20 hours). With this data, new promoter sequences could be designed and further validated by the use of RT-qPCR and reporter assays.
Project description:Mycoplasma gallisepticum is a convenient model object for studying the regulation of transcription because it has a reduced genome, lack of cell wall and many metabolic pathways, and also easy to culture and non-pathogenic to humans. For rapid investigation of gene expression we developed microarray design including 3 366 probes for 678 genes. They included 665 protein coding sequences and 13 antisense RNAs from 816 genes and 17 ncRNAs present in Mycoplasma gallisepticum. This work was carried out transcriptomic profiling for different types of effects on the expression of genes of Mycoplasma gallisepticum: 1) genetic knock-out mutants; 2) cell culture exposed to sublethal concentrations of antibiotics; and 3) well-characterized heat stress effect. The study was performed on Agilent one-color microarray with custom design and random-T7 polymerase primer for cDNA synthesis. Using set of different probes for each gene or ncRNA allows to increase accuracy of gene expression quality.
Project description:Mycoplasma hyopneumoniae causes swine pneumonia and contributes significantly to porcine respiratory disease complex. The mechanisms of pathogenesis are difficult to address since there is a lack of genetic tools, but microarrays can be used to study transcriptional changes that occur during colonization and disease in pigs. This approach has the potential to identify genes important to virulence. This study sought to identify genes that change transcript levels during infection. To accomplish this, organisms collected from bronchial alveolar lavages were compared to that of broth grown organisms. Bronchial alveolar lavage was performed on pigs 28 days post infection with M. hyopneumoniae, and organisms were isolated by differential centrifugation. Mycoplasma RNA enriched preparations were then obtained from total RNA by subtracting eucaryotic ribosomal and messenger RNAs. cDNA was generated with mycoplasma ORF-specific primers, fluorescently labeled with Cy3 and Cy5, and used to interrogate microarrays. Arrays were scanned and analyzed using a mixed linear statistical model. Nine biological replicates were analyzed in this fashion. Our analysis indicated that 33 Mhyo genes were up-regulated and 46 genes were down-regulated (p<0.01) during disease in the pig lung at a false discovery rate < 2.7%. Of the down-regulated genes, 27 of 46 (59%) lacked assigned function, and 20 of 33 (61%) of the up-regulated genes were hypothetical genes. Four down-regulated and two up-regulated genes were putative lipoproteins. secA (mhp295; p = 0.003), and two glycerol transport permeases (potA (mhp380); p = 0.006 and ugpA (mhp381); p = 0.003) were up-regulated in vivo. Elongation factor EF-G (fusA (mhp083); p = 0.002), rpoC (mhp635; p = 0.003), adenylate kinase (adk (mhp208); p = 0.001), prolyl aminoacyl tRNA synthetase (proS (mhp397); p = 0.009) and cysteinyl-tRNA synthetase (cysS (mhp661); p < 0.001) were down-regulated in vivo. Keywords: RNA, spotted DNA/cDNA