Mhyo IG Rnd vs MhRnd
Ontology highlight
ABSTRACT:
ORGANISM(S): Mesomycoplasma hyopneumoniae
PROVIDER: GSE17893 | GEO | 2010/06/30
SECONDARY ACCESSION(S): PRJNA120081
REPOSITORIES: GEO
Similar Datasets
Project description:Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia and a major factor in the porcine respiratory disease complex. A clear understanding of the mechanisms of pathogenesis does not exist. The virulence factors of M. hyopneumoniae are largely unknown and are most probably complex in nature. The transcriptional profile of intergenic regions is investigated using microarray PCR and oligonucleotide probes. It is hypothesized that intergenic regions of the Mhyo genome are being transcribed along with the ORFs. To test for transcripts in IG regions, cDNA was generated from M. hyopneumoniae RNA using random hexamers (IDT; Integrated DNA Technologies, Inc. Coralville, IA) and a set of 129 hexamer oligonucleotide primers designed for M. hyopneumoniae mRNA. A single total RNA sample was split into eight 5 μg samples, and cDNA synthesis was conducted on four of these samples using the random hexamers (IDT); for the other four samples, cDNA was generated using primers designed for M. hyopneumoniae as described by Madsen et al. 2006. In each set of four, two samples were fluorescently labeled with Cy3 and the other two were labeled with Cy5 using an indirect labeling protocol. After labeling and cleanup, all 8 samples were quantified on a Nanodrop ND-1000 spectrophotometer. For each array on the slide, 1 μg of labeled cDNA from each type of primer generation (Cy3 or Cy5, random or specific primer set) was combined and hybridized to a PCR featured array and an oligonucleotide featured array. This resulted in 2 arrays of PCR probes hybridized with dye swaps and 2 arrays of oligonucleotide probes with dye swaps. A second RNA sample was split in four 5 μg samples of total RNA, and the hybridization process was repeated for the oligonucleotide probe array.
2010-06-30 | E-GEOD-17893 | biostudies-arrayexpress
Project description:Mycoplasma hyopneumoniae, the causative agent of swine enzootic pneumonia, colonizes the cilia of swine lungs, causing ciliostasis and cell death. Mycoplasma hyopneumoniae is a component of the porcine respiratory disease complex (PRDC) and is especially problematic for the finishing swine industry, causing the loss of hundreds of millions of dollars in farm revenues worldwide. For successful infection, M. hyopneumoniae must effectively resist oxidative stresses due to the release of oxidative compounds from neutrophils and macrophages during the host’s immune response. However, the mechanism M. hyopneumoniae uses to avert the host response is still unclear. To gain a better understanding of the transcriptional responses of M. hyopneumoniae under oxidative stress, cultures were grown to early exponential phase and exposed to 0.5% percent hydrogen peroxide for 15 minutes. RNA samples from these cultures were collected and compared to RNA samples from control cultures using two-color PCR-based M. hyopneumoniae microarrays. This study revealed significant down-regulation of important glycolytic pathway genes and gene transcription proteins, as well as a protein known to activate oxidative stressor cascades in neutrophils. This study has also contained significantly differentially expressed genes common to other environmental stress responses, and merits further study of universal stress response genes of M. hyopneumoniae. Keywords: Mycoplasma hyopneumoniae, RNA microarray
2007-11-01 | GSE8304 | GEO
Project description:Mycoplasma hyopneumoniae pathogenic strains, like 7448, are the causative agents of porcine enzootic pneumonia. Non-pathogenic strains, like M. hyopneumoniae J, does not cause disease, although shares the entire repertoire of known virulence-related genes with M. hyopneumoniae 7448. In this context, the differential expression of ortholog genes is likely responsible, at least in part, for differences in pathogenicity or virulence level between these strains. Moreover, in the porcine lung, M. hyopneumoniae faces a hostile environment, with both oxidative and heat stresses. The performed comparative proteomics analyses provided evidence of differential stress responses between a pathogenic and a non-pathogenic M. hyopneumoniae strain, involving tens of proteins, including some known virulence factors. The results suggest that stress conditions trigger the expression of potential virulence factors in the pathogenic M. hyopneumoniae 7448, but not in the non-pathogenic M. hyopneumoniae J.
2019-03-29 | PXD012742 | Pride
Project description:Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia and a major factor in the porcine respiratory disease complex. A clear understanding of the mechanisms of pathogenesis does not exist although it is clear that M. hyopneumoniae adheres to porcine ciliated epithelium by action of a protein called P97. Previous studies have shown variation in the gene encoding the P97cilium adhesin within different strains of M. hyopneumoniae, but the extent of genetic variation among field strains across the genome is not known. Since M. hyopneumoniae is a worldwide problem, it is reasonable to expect that a wide range of genetic variability may exist given all of the different breed and housing conditions. This variation may impact the overall virulence of a single strain. Using microarray technology, this study examined potential variation of fourteen field strains in comparison to strain 232 on which the array was based. Genomic DNA was obtained, amplified with TempliPhi™, and labeled indirectly with Alexa dyes. Post genomic hybridization, the arrays were scanned and data analyzed using a linear statistical model. Results indicate that genetic variation could be detected in all fourteen field strains but across different loci, suggesting that variation occurs throughout the genome. Fifty-nine percent of the variable loci were hypothetical genes. Twenty-two percent of the lipoprotein genes showed variation in at least one field strain. A permutation test identified a location in M. hyopneumoniae genome where spatial clustering of variability between the field strains and strain 232 exists. Keywords: CGH, Mycoplasma Hyopneumoniae
2008-01-01 | GSE8306 | GEO
Project description:Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia and a major factor in the porcine respiratory disease complex. A clear understanding of the mechanisms of pathogenesis does not exist although it is clear that M. hyopneumoniae adheres to porcine ciliated epithelium by action of a protein called P97. Previous studies have shown variation in the gene encoding the P97cilium adhesin within different strains of M. hyopneumoniae, but the extent of genetic variation among field strains across the genome is not known. Since M. hyopneumoniae is a worldwide problem, it is reasonable to expect that a wide range of genetic variability may exist given all of the different breed and housing conditions. This variation may impact the overall virulence of a single strain. Using microarray technology, this study examined potential variation of fourteen field strains in comparison to strain 232 on which the array was based. Genomic DNA was obtained, amplified with TempliPhiâ¢, and labeled indirectly with Alexa dyes. Post genomic hybridization, the arrays were scanned and data analyzed using a linear statistical model. Results indicate that genetic variation could be detected in all fourteen field strains but across different loci, suggesting that variation occurs throughout the genome. Fifty-nine percent of the variable loci were hypothetical genes. Twenty-two percent of the lipoprotein genes showed variation in at least one field strain. A permutation test identified a location in M. hyopneumoniae genome where spatial clustering of variability between the field strains and strain 232 exists. TempliPhi⢠amplified DNA samples from field isolates were compared to control strain 232 using a two-color experimental microarray design. Independent samples from one isolate labeled with one dye were paired with control samples labeled with the alternate dye; the samples were mixed and hybridized to the microarray. For nine of the fourteen isolates (95MP1501, 95MP1502, 95MP1503, 95MP1504, 95MP1508, 95MP1509, 97MP0001, 00MP1301, and 05MP2301), four independent field isolate DNA samples were paired with four independent DNA samples from control 232. In two of the four arrays, the control sample was labeled with Alexa 555 dye and compared to the field isolate sample labeled with Alexa 647 dye (Molecular Probes, Inc., Eugene, Ore.). The dye assignment to control and treated samples was reversed for the other two arrays (dye swap). The arrays were hybridized under identical conditions as described below. This procedure was repeated for isolates 95MP1510 for a total of four arrays, where the control sample was labeled with Alexa 647 dye in three of the arrays and Alexa 555 dye for the fourth array. For isolates 95MP1505, 95MP1506 and 95MP1507, a total of five arrays each, including two dye swaps, were done; and for isolate 00MP1502, a total of six arrays were done with the control labeled with Alexa 555 dye for four of the arrays and Alexa 647 for the other two arrays.
2010-05-27 | E-GEOD-8306 | biostudies-arrayexpress
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
2008-01-01 | GSE9057 | GEO
Project description:Investigation of whole genome gene expression level changes in porcine alveolar macrophage response to co-infection of PRRSV and M. hyopneumoniae The results in this study will be further described in Bin li et al., 2013. BMC genomics. A 24 chip study using total RNA recovered from four separate group include control, PRRSV, M. hyopneumoniae and PRRSV+M. hyopneumoniae.
2014-05-30 | E-GEOD-49882 | biostudies-arrayexpress
Project description:Mycoplasma hyopneumoniae Transcriptome
| PRJNA275209 | ENA
Project description:Causes enzootic pneumonia in swine
| PRJNA12343 | ENA
Project description:Immune traits (ITs) are potentially relevant criteria to characterize individual’s immunocompetence. Thus, porcine ITs related to innate and adaptive immunity were studied by functional genomics approaches, with no initial focus on resistance to specific pathogens. Peripheral blood transcriptome was analysed in 60 days old Large White pigs (N=445) 3 weeks after vaccination against Mycoplasma hyopneumoniae. Groups of 4 to 10 animals classified in the extreme tails of CD4-/CD8+and TCRγδ+ cell counts, phagocytosis, in vitro production of IL2, IL10, TNF and IFNG, and anti-Mycoplasma antibodies distributions were selected for transcriptome studies with a porcine generic array enriched with immunity-related genes (SLA-RI/NRSP8-13K). Among the ITs studied, transcriptome analysis revealed differentially expressed genes for CD4-/CD8+cell counts, phagocytosis, and in vitro production of IL2 and IL10. A subset of these genes was confirmed by real time qPCR. Gene set enrichment analysis showed a significant over-representation of immune response functions. An independent set of 74 no overlapping animals was employed for validation and a total of 5 potential gene biomarkers were found for prediction of immunocompetence. These biomarkers performed with 79% sensitivity (95% CI 61% to 97%) and 86% specificity (95% CI 72% to 100%). Considering the observed transcriptome differences in animals with extreme ITs levels, we conclude that gene expression profiling appears promising as a tool for biological monitoring of genetic variance in pigs.
2013-12-19 | GSE45196 | GEO