EV-TRACK

Search > Results

You searched for: EV230846 (EV-TRACK ID)

Showing 1 - 1 of 1

Results list Study Tree

Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230846	1/1	Edwardsiella piscicida	EIB202	(d)(U)C DG Filtration ultrafiltration	Zhang L	2018	44%
Study summary Full title Systematic Identification of Intracellular-Translocated Candidate Effectors in . All authors Zhang L, Jiang Z, Fang S, Huang Y, Yang D, Wang Q, Zhang Y, Liu Q Journal Front Cell Infect Microbiol Abstract Many bacterial pathogens inject effectors directly into host cells to target a variety of host cellu (show more...)Many bacterial pathogens inject effectors directly into host cells to target a variety of host cellular processes and promote bacterial dissemination and survival. Identifying the bacterial effectors and elucidating their functions are central to understanding the molecular pathogenesis of these pathogens. is a pathogen with a wide host range, and very few of its effectors have been identified to date. Here, based on the genes significantly regulated by macrophage infection, we identified 25 intracellular translocation-positive candidate effectors, including all five previously reported effectors, namely EseG, EseJ, EseH, EseK, and EvpP. A subsequent secretion analysis revealed diverse secretion patterns for the 25 effector candidates, suggesting that multiple transport pathways were involved in the internalization of these candidate effectors. Further, we identified two novel type VI secretion system (T6SS) putative effectors and three outer membrane vesicles (OMV)-dependent putative effectors among the candidate effectors described above, and further analyzed their contribution to bacterial virulence in a zebrafish model. This work demonstrates an effective approach for screening bacterial effectors and expands the effectors repertoire in . (hide) EV-METRIC 44% (84th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles OMV (outer membrane vesicles) Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration ultrafiltration Protein markers EV: OmpA/ HA non-EV: RNAP Proteomics no Show all info Study aim Identification of content (omics approaches) Sample Species Edwardsiella piscicida Sample Type Cell culture supernatant EV-producing cells EIB202 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: time(min) 90 Pelleting: rotor type NVTTM65 Pelleting: speed (g) 284000 Density gradient Type Discontinuous Number of initial discontinuous layers 5 Lowest density fraction 20% Highest density fraction 45% Total gradient volume, incl. sample (mL) 11 mL Sample volume (mL) 2 mL (mi Orientation Bottom-up Rotor type NVTTM65 Speed (g) 284000 Duration (min) 960 Fraction volume (mL) 1 mL Fraction processing Centrifugation Pelleting: speed (g) 284000 Pelleting: adjusted k-factor TDB Filtration steps 0.45µm > x > 0.22µm, Ultra filtration Cut-off size (kDa) 30 Membrane type not specified Other Name other separation method ultrafiltration Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins OmpA/ HA Not detected contaminants RNAP Characterization: Lipid analysis No Characterization: Particle analysis EM EM-type Transmission-EM Image type Close-up, Wide-field

				1 - 1 of 1

EV-TRACK ID	EV230846
species	Edwardsiella piscicida
sample type	Cell culture
cell type	EIB202
condition	Control condition
separation protocol	dUC/ Density gradient/ Filtration/ ultrafiltration
Exp. nr.	1
EV-METRIC %	44