Mariprofundus ferroooxydans PV-1 osmotic shock vs extraction with NaOH LC-MSMS
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ABSTRACT: Proteomic analysis was performed on protein extracts of M. ferrooxydans by following two methodologies: osmotic shock and alkalyne lysis (extraction with NaOH).
Project description:Pseudomonas syringae pv. tomato DC3000 (Pst) is a virulent pathogen, which causes disease on tomato and Arabidopsis. The type III secretion system (TTSS) plays a key role in pathogenesis by translocating virulence effectors from the bacteria into the plant host cell, while the phytotoxin coronatine (COR) contributes to virulence and disease symptom development. Recent studies suggest that both the TTSS and and COR are involved in the suppression of host basal defenses. However, little is known about the interplay between the host gene expression associated with basal defenses and the virulence activities of the TTSS and COR during infection. The global effects of the TTSS and COR on host gene expression associated with other host cellular processes during bacterial infection are also not well characterized. In this study, we used the Affymetrix full genome chip to determine the Arabidopsis transcriptome associated with basal defense to Pst DC3000 hrp mutants and the human pathogenic bacterium Escherichia coli O157:H7. We then used Pst DC3000 virulence mutants to characterize Arabidopsis transcriptional responses to the action of hrp-regulated virulence factors (e.g., TTSS and COR) during bacterial infection. Additionally, we used bacterial fliC mutants to assess the role of the PAMP flagellin in induction of basal defense-associated transcriptional responses. In total, our global gene expression analysis identified more than 5000 Arabidopsis genes that are reproducibly regulated more than 2-fold in three independent biological replicates of at least one type of comparison. Regulation of these genes provides a molecular signature for Arabidopsis basal defense to plant and human pathogenic bacteria, and illustrates both common and distinct global virulence effects of the TTSS, COR, and possibly other hrp-regulated virulence factors during Pst DC3000 infection. Experimenter name = William Underwood; Experimenter phone = 517-353-9182; Experimenter fax = 517-353-9168; Experimenter address = Michigan State University; Experimenter address = 222 Plant Biology Building; Experimenter address = 178 Wilson R.d. Experimenter address = East Lansing, MI; Experimenter zip/postal_code = 48824; Experimenter country = USA Experiment Overall Design: 40 samples were used in this experiment
Project description:A proteomic analysis of a redox active membrane fraction (doublet at >200 kDa) of Mariprofundus ferrooxydans, a chemolithoautotrophic, neutrophilic, iron-oxidizing Zetaproteobacterium.
Project description:The protozoan Giardia lamblia is a unique biological model to investigate minimal cellular mechanisms because it is an early‐diverged eukaryote that has undergone reductive evolution and has minimized or lost organelles such as mitochondria, peroxisomes, the endo-lysosomal system and a Golgi apparatus. Giardia trophozoites reside in the host intestines where they largely depend on scavenging metabolites from the surrounding through endocytosis and/or pinocytosis. A canonical endo‐lysosomal system however is inexistent, but small peripheral vesicles (PVs) have been described which have partial endocytic characteristics but lack endocytic hallmarks: (i) unlike the dynamic multivesicular endo-lysosomal system in mammalian cells, PVs have a fixed location at the cell periphery and show no lateral cargo exchange; (ii) PVs lack maturation-specific markers. For transmission to a new host, motile trophozoites differentiate to cysts and thus secrete a protective cyst wall (CW). Giardia however has no Golgi or to organize regulated secretion of CW material. Instead, differentiating cells generate specialized organelles termed encystation‐specific vesicles (ESVs), being stage-regulated Golgi relic organelles dedicated to post‐translational maturation of the CW material. Basic sorting processes in maturing ESVs have been identified recently, but underlying mechanisms remain unknown. To define the first proteome of ESVs and PVs, we used a new strategy combining flow cytometry-based organelle sorting with in silico filtration of mass spectrometry data.
Project description:The transcriptomic modulations leading to defense response in rice one hour after inoculation by Xanthomonas oryzae pv oryzae. Xoo and mock inoculated plant of cultivars IET8585 (bacterial leaf blight resistant) and IR-24 (bacterial leaf blight susceptible) were compared.
Project description:Adenocarcinoma cell line SW620 were treated with tetradecylthioacetic acid,TTA, or control (NaOH) for 24 h. degrees RNA isolated, Affymetrix gene expression
Project description:This experiment analyses the expression data of the wild type P. syringae pv. tomato DC3000 grown in the absence and in the presence of phloretin and naringenin.
Project description:P. syringae pv. phaseolicola is the causal agent of the halo blight disease of beans (Phaseolus vulgaris L). The disease attacks both foliage and pods of plant host. Many genes involve in pathogenicity and virulence are induced only in plant or in the presence of host components. In this work we investigated the effect of bean pod extract on the transcriptomic profile of the bacterium, when grown at low temperature in minimal medium with or without bean pod extract. Two RNA samples were compared, one prepared from cells grown in minimal medium M9 and the other from cells grown in minimal medium supplemented with bean pod extract.To control de biological variation that might interfere with data interpretation, a minimum of three biological replicates and two technical replicates (swap) were prepared.
Project description:Transcription profiling of the DSF regulon in Xanthomonas oryzae pv. oryzae (Xoo) using wild type and the rpfF mutant. Cell-cell signaling mediated by the quorum sensing molecule known as Diffusible Signaling factor (DSF) is required for virulence of Xanthomonas group of plant pathogens. DSF in different Xanthomonas and the closely related plant pathogen Xylella fastidiosa regulates diverse traits in a strain specific manner. The transcriptional profiling performed in this study is to elucidate the traits regulated by DSF from the Indian isolate of Xanthomonas oryzae pv. oryzae, which exhibits traits very different from other Xanthomonas group of plant pathogen. In this study, transcription analysis was done between a wild type Xanthomonas oryzae pv. oryzae strain and an isogenic strain that has a mutation in the DSF biosynthetic gene rpfF. Agilent one-color experiment, Organism: Xanthomonas oryzae, Agilent-025096 Genotypic Technology Pvt. Ltd. designed Custom Xanthomonas oryzae 8x15k, Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442).
Project description:This animal study was approved by the Ethics Committee at school of Chinese medicine,Hong Kong Baptist University. A total of 4 female and 4 male C57BL/6 mices were included in control diet group (BC); A total of 4 female and 4 male C57BL/6 mices were included in high fat diet group (BT).