Dataset Information

Label-free quantitative proteomic analysis of Brucella abortus and Brucella melitensis

ABSTRACT: Brucella type strains (B. abortus 544 and B. melitensis 16 M), field-isolates (B. abortus T and B. melitensis C) were independently cultivated six times on Tryptic Soy Broth at 37 °C in the presence of 5% CO2 until mid-logarithmic phase. The cells were harvested by brief centrifugation and the bacteria was inactivated using recommended ethanol precipitation procedure. Following which the cells were lysed, proteins quantified and the differentially expressed proteins among B. abortus and B. melitensis were identified using liquid chromatography-mass spectrometry (LC/MS) based label free quantitative proteomics analysis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Brucella Abortus Brucella Melitensis

DISEASE(S): Brucella Melitensis Brucellosis,Brucellosis,Brucella Abortus Brucellosis

SUBMITTER: Jayaseelan Murugaiyan

LAB HEAD: Jayaseelan Murugaiyan

PROVIDER: PXD006348 | Pride | 2020-06-11

REPOSITORIES: pride

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Publications

Pan-Proteomic Analysis and Elucidation of Protein Abundance among the Closely Related Brucella Species, Brucella abortus and Brucella melitensis.

Murugaiyan Jayaseelan J Eravci Murat M Weise Christoph C Roesler Uwe U Sprague Lisa D LD Neubauer Heinrich H Wareth Gamal G

Biomolecules 20200530 6

Brucellosis is a zoonotic infection caused by bacteria of the genus Brucella. The species, B. abortus and B. melitensis, major causative agents of human brucellosis, share remarkably similar genomes, but they differ in their natural hosts, phenotype, antigenic, immunogenic, proteomic and metabolomic properties. In the present study, label-free quantitative proteomic analysis was applied to investigate protein expression level differences. Type strains and field strains were ...[more]

PMID: 32486122

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Project description:Brucella spp. is an intracellular pathogen in vivo. The intracellular B. melitensis transcriptome was determined by initially enriched and then amplified B. melitensis RNA from total RNA of B. melitensis-infected HeLa cells. Analysis of microarray results identified 161 and 115 genes differentially expressed at 4 and 12 h p.i., respectively. Most of the genes (78%) differentially expressed were down-regulated at the earliest time point, but up-regulated (75%) at 12 h p.i. The analysis of the results indicates that Brucella undergo an adaptation period during the first 4 h p.i. that is overcome by 12 h p.i., permitting Brucella to replicate intracellularly. Specific genes and biological processes identified in this study will further help elucidate how Brucella act during the early infectious process to their eventual benefit and to the detriment of the naM-CM-/ve host. Keywords: Time course study of intracellular B. melitensis gene expression Gene expression of the intracellular Brucella melitensis was determined at 4 and 12 h p.i. We generated the following samples: A) B. melitensis total RNA enriched and amplified from total RNA of B. melitensis-infected HeLa cells at 4 h p.i.; B) Total RNA isolated from B. melitensis-infected HeLa cells at 4 h p.i.; C) B. melitensis total RNA enriched and amplified from total RNA of B. melitensis-infected HeLa cells at 12 h p.i.; D) Total RNA isolated from B. melitensis-infected HeLa cells at 12 h p.i. B. melitensis total RNA was initially enriched and then amplified from total RNA of B. melitensis-infected HeLa cells at 4 and 12 h p.i. in quadruplicate, indirectly labeled and co-hybridized against B. melitensis gDNA to a custom 3.2K B. melitensis oligo-array (n = 8). As there was a possibility that some HeLa transcripts cross-hybridize with probes on B. melitensis microarrays, the original total RNA from B. melitensis-infected HeLa cells were also co-hybridized against B. melitensis gDNA to B. melitensis oligo-arrays (n = 8), and any oligospots with signals were considered non-specific and eliminated from all analysis to avoid false positive gene detection. The intracellular B. melitensis gene expression was compared to the gene expression of the inoculum (n = 2). Every Brucella melitensis open reading frame was printed in triplicate on each microarray, thereby providing three technical replicates for each biological replicates. Each replicate was normalized against labeled Brucella melitensis genomic DNA.

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Dataset Information

Label-free quantitative proteomic analysis of Brucella abortus and Brucella melitensis

Dataset's files

Publications

Pan-Proteomic Analysis and Elucidation of Protein Abundance among the Closely Related <i>Brucella</i> Species, <i>Brucella abortus</i> and <i>Brucella melitensis</i>.

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