sars cov nsp3 protein (Sino Biological)


Name:
SARS CoV 2 2019 nCoV NSP3 His Recombinant Protein
Description:
A DNA sequence encoding the SARS CoV 2 2019 nCoV NSP3 YP 009725299 1 Ile205 Lys379 was expressed with a polyhistidine tag at the C terminus
Catalog Number:
40638-V07E
Price:
None
Category:
recombinant protein
Host:
Baculovirus-Insect Cells
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Structured Review
![<t>CoV</t> RNA synthesis is confined to RO regions. Newly synthesized vRNA was metabolically labeled by providing tritiated uridine to CoV-infected cells pretreated with actinomycin D to limit host transcription. (A) Analysis of the amount of radioactive label incorporated into RNA as a function of the labeling time in <t>SARS-CoV-infected</t> Vero E6 cells (MOI 10), as measured by scintillation counting on the RNA isolated from the cells (underlying numerical data in S2 Data ). The label was provided simultaneously to all the samples at 6 hpi. (B-D) EM detection by autoradiography. (B) Overview of a SARS-CoV-infected Vero E6 cell (MOI 10, 7 hpi, labeled for 20 minutes). Autoradiography grains accumulate in the RO regions. Scale bar, 1 μm. (C, D) Quantification of the autoradiography signal per subcellular structure (see also S3 Data ). Labeling densities and RLIs in different subcellular regions of (C) Vero E6 cells infected with SARS-CoV (MOI 10) or (D) Huh7 cells infected with MERS-CoV (MOI 5). Radioactively labeled uridine was provided for the indicated periods of time immediately before fixation at 7 hpi and 12 hpi, respectively. These time points represent, respectively, the middle (SARS-CoV) or late (MERS-CoV) exponential phase of viral replication [ 21 , 34 ]. Control mock-infected cells are excluded from the RLI plots, as RLI comparisons between conditions require the same number of classes (subcellular regions) and these cells lack ROs and virions. CM, convoluted membranes; CoV, coronavirus; cpm, counts per minute; DMS, double-membrane spherule; DMV, double-membrane vesicle; EM, electron microscopy; ER, endoplasmic reticulum; HCoV-229E, human coronavirus 229E; hpi, hours postinfection; IBV, infectious bronchitis virus; LD, lipid droplet; m, mitochondrion; MERS-CoV, Middle East respiratory syndrome-CoV; MHV, murine hepatitis virus; MOI, multiplicity of infection; N, nucleus; RLI, relative labeling index;; RO, replication organelle; SARS-CoV, severe acute respiratory syndrome-CoV; VCR, virion-containing region; vRNA, viral RNA.](https://storage.googleapis.com/bioz_article_images/PMC7302735/pbio.3000715.g004.jpg)
A DNA sequence encoding the SARS CoV 2 2019 nCoV NSP3 YP 009725299 1 Ile205 Lys379 was expressed with a polyhistidine tag at the C terminus
https://www.bioz.com/result/sars cov nsp3 protein/product/Sino Biological
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Images
1) Product Images from "A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis"
Article Title: A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis
Journal: PLoS Biology
doi: 10.1371/journal.pbio.3000715
![... as a function of the labeling time in SARS-CoV-infected Vero E6 cells (MOI 10), as measured by ... CoV RNA synthesis is confined to RO regions. Newly synthesized vRNA was metabolically labeled by providing tritiated uridine to CoV-infected cells pretreated with actinomycin D to limit host transcription. (A) Analysis of the amount of radioactive label incorporated into RNA as a function of the labeling time in SARS-CoV-infected Vero E6 cells (MOI 10), as measured by scintillation counting on the RNA isolated from the cells (underlying numerical data in S2 Data ). The label was provided simultaneously to all the samples at 6 hpi. (B-D) EM detection by autoradiography. (B) Overview of a SARS-CoV-infected Vero E6 cell (MOI 10, 7 hpi, labeled for 20 minutes). Autoradiography grains accumulate in the RO regions. Scale bar, 1 μm. (C, D) Quantification of the autoradiography signal per subcellular structure (see also S3 Data ). Labeling densities and RLIs in different subcellular regions of (C) Vero E6 cells infected with SARS-CoV (MOI 10) or (D) Huh7 cells infected with MERS-CoV (MOI 5). Radioactively labeled uridine was provided for the indicated periods of time immediately before fixation at 7 hpi and 12 hpi, respectively. These time points represent, respectively, the middle (SARS-CoV) or late (MERS-CoV) exponential phase of viral replication [ 21 , 34 ]. Control mock-infected cells are excluded from the RLI plots, as RLI comparisons between conditions require the same number of classes (subcellular regions) and these cells lack ROs and virions. CM, convoluted membranes; CoV, coronavirus; cpm, counts per minute; DMS, double-membrane spherule; DMV, double-membrane vesicle; EM, electron microscopy; ER, endoplasmic reticulum; HCoV-229E, human coronavirus 229E; hpi, hours postinfection; IBV, infectious bronchitis virus; LD, lipid droplet; m, mitochondrion; MERS-CoV, Middle East respiratory syndrome-CoV; MHV, murine hepatitis virus; MOI, multiplicity of infection; N, nucleus; RLI, relative labeling index;; RO, replication organelle; SARS-CoV, severe acute respiratory syndrome-CoV; VCR, virion-containing region; vRNA, viral RNA.](https://storage.googleapis.com/bioz_article_images/PMC7302735/pbio.3000715.g004.jpg)
Figure Legend Snippet: CoV RNA synthesis is confined to RO regions. Newly synthesized vRNA was metabolically labeled by providing tritiated uridine to CoV-infected cells pretreated with actinomycin D to limit host transcription. (A) Analysis of the amount of radioactive label incorporated into RNA as a function of the labeling time in SARS-CoV-infected Vero E6 cells (MOI 10), as measured by scintillation counting on the RNA isolated from the cells (underlying numerical data in S2 Data ). The label was provided simultaneously to all the samples at 6 hpi. (B-D) EM detection by autoradiography. (B) Overview of a SARS-CoV-infected Vero E6 cell (MOI 10, 7 hpi, labeled for 20 minutes). Autoradiography grains accumulate in the RO regions. Scale bar, 1 μm. (C, D) Quantification of the autoradiography signal per subcellular structure (see also S3 Data ). Labeling densities and RLIs in different subcellular regions of (C) Vero E6 cells infected with SARS-CoV (MOI 10) or (D) Huh7 cells infected with MERS-CoV (MOI 5). Radioactively labeled uridine was provided for the indicated periods of time immediately before fixation at 7 hpi and 12 hpi, respectively. These time points represent, respectively, the middle (SARS-CoV) or late (MERS-CoV) exponential phase of viral replication [ 21 , 34 ]. Control mock-infected cells are excluded from the RLI plots, as RLI comparisons between conditions require the same number of classes (subcellular regions) and these cells lack ROs and virions. CM, convoluted membranes; CoV, coronavirus; cpm, counts per minute; DMS, double-membrane spherule; DMV, double-membrane vesicle; EM, electron microscopy; ER, endoplasmic reticulum; HCoV-229E, human coronavirus 229E; hpi, hours postinfection; IBV, infectious bronchitis virus; LD, lipid droplet; m, mitochondrion; MERS-CoV, Middle East respiratory syndrome-CoV; MHV, murine hepatitis virus; MOI, multiplicity of infection; N, nucleus; RLI, relative labeling index;; RO, replication organelle; SARS-CoV, severe acute respiratory syndrome-CoV; VCR, virion-containing region; vRNA, viral RNA.
Techniques Used: Synthesized, Metabolic Labelling, Labeling, Infection, Isolation, Autoradiography, Electron Microscopy

Figure Legend Snippet: IEM detection of viral markers in MERS-CoV-infected cells. (A-G) Immunogold labeling of thawed cryo-sections of MERS-CoV-infected Huh7 cells (12 hpi) for the detection of the indicated viral proteins. (A-C) Structural proteins were detected on virions (black arrowheads) and, for the M and S proteins, also on Golgi cisterna. While regions containing DMS (white arrowheads) and CM labeled for the N protein (D) and nsp3 (G), the M and S protein were not detected in these areas. (H-I) Immunogold labeling of dsRNA in HPF-FS samples of MERS-CoV-infected Huh7 cells (13 hpi). The label accumulated on DMVs, which could be easily detected in this type of samples (black arrows), whereas the regions with CM and DMSs, which appeared as dark areas among the DMV clusters, were devoid of dsRNA signal. Scale bars, 250 nm. CM, convoluted membranes; DMS, double-membrane spherule; dsRNA, double-stranded RNA; G, Golgi complex; HPF-FS, high-pressure freezing, freeze-substitution; hpi, hours postinfection; IEM, immunoelectron microscopy; m, mitochondrion; MERS-CoV, Middle East respiratory syndrome-CoV; nsp3, nonstructural protein 3.
Techniques Used: Infection, Labeling, Immuno-Electron Microscopy
![... mammal cells infected with (from left to right) SARS-CoV (MOI 10, 9 hpi), MHV (MOI 10, 8 ... DMSs are induced by diverse beta- and alpha-CoVs. 2D-EM images from 100-nm-thick sections of different mammal cells infected with (from left to right) SARS-CoV (MOI 10, 9 hpi), MHV (MOI 10, 8 hpi), and HCoV-229E (MOI 5, 24 hpi). These time points represent intermediate to late stages in infection [ 34 – 36 ]. Both beta-CoVs (A,B) and the alpha-CoV (C) induce membrane modifications that include not only DMVs (asterisks) and CM but also DMSs (white arrowheads). Scale bars, 250 nm. CM, convoluted membranes; CoV, coronavirus; DMS, double-membrane spherule; DMV, double-membrane vesicle; EM, electron microscopy; ER, endoplasmic reticulum; HCoV-229E, human coronavirus 229E; hpi, hours postinfection; IBV, infectious bronchitis virus; MHV, murine hepatitis virus; MOI, multiplicity of infection; SARS-CoV, severe acute respiratory syndrome-CoV.](https://storage.googleapis.com/bioz_article_images/PMC7302735/pbio.3000715.g003.jpg)
Figure Legend Snippet: DMSs are induced by diverse beta- and alpha-CoVs. 2D-EM images from 100-nm-thick sections of different mammal cells infected with (from left to right) SARS-CoV (MOI 10, 9 hpi), MHV (MOI 10, 8 hpi), and HCoV-229E (MOI 5, 24 hpi). These time points represent intermediate to late stages in infection [ 34 – 36 ]. Both beta-CoVs (A,B) and the alpha-CoV (C) induce membrane modifications that include not only DMVs (asterisks) and CM but also DMSs (white arrowheads). Scale bars, 250 nm. CM, convoluted membranes; CoV, coronavirus; DMS, double-membrane spherule; DMV, double-membrane vesicle; EM, electron microscopy; ER, endoplasmic reticulum; HCoV-229E, human coronavirus 229E; hpi, hours postinfection; IBV, infectious bronchitis virus; MHV, murine hepatitis virus; MOI, multiplicity of infection; SARS-CoV, severe acute respiratory syndrome-CoV.
Techniques Used: Infection, Electron Microscopy
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