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.
2021-05-25 | GSE152865 | GEO
Project description:Whole-genome sequencing of Mycoplasma bovis isolates
Project description:This study was aimed to elucidate a global antigenic profile of Mycoplasma bovis (M. bovis) with immunoproteomics, immunoinformatics, and gene expression approaches. The extracts of whole-cell proteins and TX-114 membrane fraction of a Chinese strain M. bovis HB0801 were separated with two dimensional gel electrophoresis (2-DE) and proteins reacting with antisera to M. bovis from experimentally infected calves were detected by MALDI-TOF MS.
Project description:Mycoplasmas are simple, but successful parasites that have the smallest genome of any free-living cell and are thought to have a highly streamlined cellular metabolism. Here we have undertaken a detailed metabolomic analysis of two species, Mycoplasma bovis and Mycoplasma gallisepticum, which cause economically important diseases in cattle and poultry, respectively. Untargeted GC/MS and LC/MS analyses of mycoplasma metabolite extracts revealed significant differences in the steady state levels of many metabolites in central carbon metabolism, while 13C stable isotope labelling studies revealed marked differences in carbon source utilization. These data were mapped onto in silico metabolic networks predicted from genome wide annotations. The analyses elucidated distinct differences, including a clear difference in glucose utilisation, with a marked decrease in glucose uptake and glycolysis in M. bovis compared to M. gallisepticum, which may reflect differing host nutrient availabilities. The 13C-labeling patterns also revealed several functional metabolic pathways that were previously unannotated in these species, allowing us to assign putative enzyme functions to the products of a number of genes of unknown function, especially in M. bovis. This study demonstrates the considerable potential of metabolomic analyses to assist in characterising significant differences in the metabolism of different bacterial species, and in improving genome annotation.
Project description:In past, resistance mechanisms have been identified by analysis of resistant isolates or defined mutants. Recently, high-throughput transposon mutagenesis coupled with sequencing (TraDIS-Xpress) is another approach proving useful for elucidating the roles of genes involved in the overall cellular response to a particular stress. In this study, we used TraDIS-Xpress to determine the role played by genes following exposure to colistin stress. Approximately 10^7 cells from the mutant library were inoculated into LB broth at a range of doubling concentrations of colistin ( 0.25 x MIC, 0.5 x MIC, 1 x MIC, 2 X MIC). Experiments were performed with no induction, or with induction using 0.2 or 1 mM of Isopropyl β-D-1-thiogalactopyranoside (IPTG). All experiments were performed in duplicate.
Project description:Cyclic di-AMP is an essential and ubiquitous second messenger that regulates bacterial potassium (K+) concentrations to maintain osmotic equilibrium. And its specific K+ regulation mechanism in Mycoplasma has rarely been investigated. We used the ruminant pathogen Mycoplasma bovis (M. bovis) to investigate this mechanism. We verified that MbovP496 is a c-di-AMP synthetase and its mutant T5.415 showed growth inhibition under high K+ condition. To deliver why the mutant strain T5.415 can regulate M. bovis growth, we held a RNA-seq analysis to reveal that the differentially expressed genes between WT and T5.415 under normal condition and high K+ condition. The K+ transport pathways and metabolism related pathways were enriched. In conclusion, we found that the mutation of c-di-AMP synthetase, MbovP496 can broadly influence not only the metal ions transport pathway, but also has an effect on metabolism pathways,which could significantly contribute to understanding the regulation of growth by c-di-AMP synthetase in mycoplasmas.