sars cov 2 2019 ncov nucleocapsid orf mammalian expression plasmid codon optimized covid 19 nucleocapsid research  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Nucleocapsid ORF mammalian expression plasmid Codon Optimized COVID 19 Nucleocapsid Research
    Description:
    Full length Clone DNA of SARS CoV 2 2019 nCoV Nucleoprotein NP
    Catalog Number:
    vg40588-ut
    Product Aliases:
    coronavirus NP cDNA ORF Clone 2019-nCoV, coronavirus Nucleocapsid cDNA ORF Clone 2019-nCoV, coronavirus Nucleoprotein cDNA ORF Clone 2019-nCoV, cov np cDNA ORF Clone 2019-nCoV, ncov NP cDNA ORF Clone 2019-nCoV, NCP-CoV Nucleocapsid cDNA ORF Clone 2019-nCoV, novel coronavirus NP cDNA ORF Clone 2019-nCoV, novel coronavirus Nucleocapsid cDNA ORF Clone 2019-nCoV, novel coronavirus Nucleoprotein cDNA ORF Clone 2019-nCoV, np cDNA ORF Clone 2019-nCoV, nucleocapsid cDNA ORF Clone 2019-nCoV, Nucleoprotein cDNA ORF Clone 2019-nCoV
    Price:
    195.0
    Applications:
    Stable or Transient mammalian expression
    Size:
    1Unit
    Category:
    cDNA Clone
    Molecule Name:
    NP-CoV
    Buy from Supplier


    Structured Review

    Sino Biological sars cov 2 2019 ncov nucleocapsid orf mammalian expression plasmid codon optimized covid 19 nucleocapsid research
    SARS CoV 2 2019 nCoV Nucleocapsid ORF mammalian expression plasmid Codon Optimized COVID 19 Nucleocapsid Research
    Full length Clone DNA of SARS CoV 2 2019 nCoV Nucleoprotein NP
    https://www.bioz.com/result/sars cov 2 2019 ncov nucleocapsid orf mammalian expression plasmid codon optimized covid 19 nucleocapsid research/product/Sino Biological
    Average 93 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov nucleocapsid orf mammalian expression plasmid codon optimized covid 19 nucleocapsid research - by Bioz Stars, 2021-02
    93/100 stars

    Images

    1) Product Images from "A single dose of recombinant VSV-∆G-spike vaccine provides protection against SARS-CoV-2 challenge"

    Article Title: A single dose of recombinant VSV-∆G-spike vaccine provides protection against SARS-CoV-2 challenge

    Journal: Nature Communications

    doi: 10.1038/s41467-020-20228-7

    Antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2. a Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with COVID-19 human convalescent serum. Representative images of five experiments are presented. Scale bars: 50 µm. b Correlation analysis of neutralization of rVSV-∆G-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each serum sample ( n = 12), NT 50 values were determined for neutralization of rVSV-∆G-spike or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between the neutralization assays. Spearman’s correlation r and p values are indicated. Source data are provided as a Source Data file.
    Figure Legend Snippet: Antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2. a Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with COVID-19 human convalescent serum. Representative images of five experiments are presented. Scale bars: 50 µm. b Correlation analysis of neutralization of rVSV-∆G-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each serum sample ( n = 12), NT 50 values were determined for neutralization of rVSV-∆G-spike or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between the neutralization assays. Spearman’s correlation r and p values are indicated. Source data are provided as a Source Data file.

    Techniques Used: Infection, Staining, Neutralization

    2) Product Images from "A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge"

    Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

    Journal: bioRxiv

    doi: 10.1101/2020.06.18.160655

    Surface antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2: (A) Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with serum from COVID-19 human convalescent serum (right panel). (B) Correlation analysis of neutralization of rVSV-ΔG-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each sera (n=12), NT 50 values were determined for neutralization of rVSV-ΔG-spike, or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between th neutralization assays. R 2 =0.911.
    Figure Legend Snippet: Surface antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2: (A) Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with serum from COVID-19 human convalescent serum (right panel). (B) Correlation analysis of neutralization of rVSV-ΔG-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each sera (n=12), NT 50 values were determined for neutralization of rVSV-ΔG-spike, or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between th neutralization assays. R 2 =0.911.

    Techniques Used: Infection, Staining, Neutralization

    Related Articles

    Polymerase Chain Reaction:

    Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), pre-cut by the same enzymes to remove the VSV-G gene.

    Article Title: A single dose of recombinant VSV-∆G-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full-length human codon-optimized S gene from pCMV3-SARS-CoV-2 S expression plasmid (Sino Biological, Cat# VG40588-UT) using the following primers: forward –atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), precut by the same enzymes to remove the VSV-G gene.

    Amplification:

    Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), pre-cut by the same enzymes to remove the VSV-G gene.

    Article Title: A single dose of recombinant VSV-∆G-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full-length human codon-optimized S gene from pCMV3-SARS-CoV-2 S expression plasmid (Sino Biological, Cat# VG40588-UT) using the following primers: forward –atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), precut by the same enzymes to remove the VSV-G gene.

    Plasmid Preparation:

    Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), pre-cut by the same enzymes to remove the VSV-G gene.

    Article Title: A single dose of recombinant VSV-∆G-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full-length human codon-optimized S gene from pCMV3-SARS-CoV-2 S expression plasmid (Sino Biological, Cat# VG40588-UT) using the following primers: forward –atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), precut by the same enzymes to remove the VSV-G gene.

    Expressing:

    Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), pre-cut by the same enzymes to remove the VSV-G gene.

    Article Title: A single dose of recombinant VSV-∆G-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full-length human codon-optimized S gene from pCMV3-SARS-CoV-2 S expression plasmid (Sino Biological, Cat# VG40588-UT) using the following primers: forward –atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), precut by the same enzymes to remove the VSV-G gene.

    Construct:

    Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), pre-cut by the same enzymes to remove the VSV-G gene.

    Article Title: A single dose of recombinant VSV-∆G-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: .. Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full-length human codon-optimized S gene from pCMV3-SARS-CoV-2 S expression plasmid (Sino Biological, Cat# VG40588-UT) using the following primers: forward –atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC. .. The amplified PCR product was digested by MluI and NheI restriction enzymes (NEB), and was ligated into the pVSV-FL+(2) vector (Kerafast), precut by the same enzymes to remove the VSV-G gene.

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    Sino Biological sars cov 2
    Structural conservation of SARS-CoV RBD. RBD is shown as colored surface. ACE2 is shown as gray cartoon. The three surface mutation sites (i.e. N354D, D364Y, and V367F) observed in <t>SARS-CoV-2</t> RBD are labeled. Mutation F342L is buried and not shown here.
    Sars Cov 2, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 2 article reviews
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    Sino Biological sars cov 2 2019 ncov nucleocapsid orf mammalian expression plasmid codon optimized covid 19 nucleocapsid research
    Antigenic similarity of rVSV-ΔG-spike and <t>SARS-CoV-2.</t> a Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with COVID-19 human convalescent serum. Representative images of five experiments are presented. Scale bars: 50 µm. b Correlation analysis of neutralization of rVSV-∆G-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each serum sample ( n = 12), NT 50 values were determined for neutralization of rVSV-∆G-spike or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between the neutralization assays. Spearman’s correlation r and p values are indicated. Source data are provided as a Source Data file.
    Sars Cov 2 2019 Ncov Nucleocapsid Orf Mammalian Expression Plasmid Codon Optimized Covid 19 Nucleocapsid Research, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 2019 ncov nucleocapsid orf mammalian expression plasmid codon optimized covid 19 nucleocapsid research/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Structural conservation of SARS-CoV RBD. RBD is shown as colored surface. ACE2 is shown as gray cartoon. The three surface mutation sites (i.e. N354D, D364Y, and V367F) observed in SARS-CoV-2 RBD are labeled. Mutation F342L is buried and not shown here.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Structural conservation of SARS-CoV RBD. RBD is shown as colored surface. ACE2 is shown as gray cartoon. The three surface mutation sites (i.e. N354D, D364Y, and V367F) observed in SARS-CoV-2 RBD are labeled. Mutation F342L is buried and not shown here.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Mutagenesis, Labeling

    Structure similarity between SARS-CoV-2 RBD and SARS-CoV RBD. RBD is shown in a space-filled model with colored surface. ACE2 is shown as gray tube model. The three glycosylation sites in SARS-CoV are labeled. Note that N 357 ST in SARS-CoV is changed to N 370 SA in SARS-CoV-2, which is different from the NXS/T pattern required for glycosylation, and hence this site is more likely to be unglycosylated. The two possible cross-reactive regions are marked with yellow circles.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Structure similarity between SARS-CoV-2 RBD and SARS-CoV RBD. RBD is shown in a space-filled model with colored surface. ACE2 is shown as gray tube model. The three glycosylation sites in SARS-CoV are labeled. Note that N 357 ST in SARS-CoV is changed to N 370 SA in SARS-CoV-2, which is different from the NXS/T pattern required for glycosylation, and hence this site is more likely to be unglycosylated. The two possible cross-reactive regions are marked with yellow circles.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Labeling

    Cross-reactivity and neutralization efficiency of SARS nAbs against SARS-CoV-2. A. Binding of SARS nAbs to SARS-CoV S1 protein were tested by ELISA. Recombinant S1 protein of SARS-CoV were coated on plates, serial diluted nAbs were added for binding to recombinant S1 protein. B. Binding of SARS nAbs to SARS-CoV-2 S1 protein were tested by ELSIA. Recombinant S1 protein of SARS-CoV-2 were coated on plates, serial diluted nAbs were added for binding to recombinant S1 protein. C. Neutralization of SARS-CoV nAbs against SARS-CoV-2 PSV. D. Antibody competition with SARS-CoV RBD binding to ACE2. Recombinant SARS-CoV RBD protein was coated on plates, nAbs and recombinant ACE2 were then added for RBD binding competition measurements.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Cross-reactivity and neutralization efficiency of SARS nAbs against SARS-CoV-2. A. Binding of SARS nAbs to SARS-CoV S1 protein were tested by ELISA. Recombinant S1 protein of SARS-CoV were coated on plates, serial diluted nAbs were added for binding to recombinant S1 protein. B. Binding of SARS nAbs to SARS-CoV-2 S1 protein were tested by ELSIA. Recombinant S1 protein of SARS-CoV-2 were coated on plates, serial diluted nAbs were added for binding to recombinant S1 protein. C. Neutralization of SARS-CoV nAbs against SARS-CoV-2 PSV. D. Antibody competition with SARS-CoV RBD binding to ACE2. Recombinant SARS-CoV RBD protein was coated on plates, nAbs and recombinant ACE2 were then added for RBD binding competition measurements.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Neutralization, Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant

    Sequence analysis and structure modeling of SARS-CoV-2 RBD and SARS-CoV RBD and their interactions with ACE2. A. RBD sequence alignment of SARS-CoV and SARS-CoV-2, highlighting the predominant residues that contribute to the interactions with ACE2. The distinct interactions of RBD and ACE2 for the two viruses are indicated by the down-pointing orange triangles and up-pointing red triangles, respectively. RBM residues are underlined. The one-residue insertion is indicated by the red arrow. Asterisks indicate positions of fully conserved residues. Colons indicate positions of strictly conserved residues. Periods indicate positions of weakly conserved residues. B. Conformational comparison between the RBD-ACE2 complex structures for SARS-CoV-2 and SARS-CoV. The RBD and ACE2 structures in the SARS-CoV-2 RBD-ACE2 complex model are shown as orange and pink tubes, respectively. The RBD and ACE2 structures in the optimized SARS-CoV RBD-ACE2 complex structure are shown as blue and green tubes, respectively. The location of noticeable subtle conformational difference is indicated by an arrow. C. Distinct interaction patterns in the SARS-CoV-2 and SARS-CoV RBD-ACE2 interfaces. Structures of RBD and ACE2 are shown as cartoon in pink and green colors, respectively. The side chains of the residues in both protein components, representing their unique interactions, are shown as sticks. Polar interactions (salt-bridge and hydrogen bond) are shown as blue dash line. Non-polar interactions (π-stack, π-anion, and hydrophobic interactions) are shown as orange dash line.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Sequence analysis and structure modeling of SARS-CoV-2 RBD and SARS-CoV RBD and their interactions with ACE2. A. RBD sequence alignment of SARS-CoV and SARS-CoV-2, highlighting the predominant residues that contribute to the interactions with ACE2. The distinct interactions of RBD and ACE2 for the two viruses are indicated by the down-pointing orange triangles and up-pointing red triangles, respectively. RBM residues are underlined. The one-residue insertion is indicated by the red arrow. Asterisks indicate positions of fully conserved residues. Colons indicate positions of strictly conserved residues. Periods indicate positions of weakly conserved residues. B. Conformational comparison between the RBD-ACE2 complex structures for SARS-CoV-2 and SARS-CoV. The RBD and ACE2 structures in the SARS-CoV-2 RBD-ACE2 complex model are shown as orange and pink tubes, respectively. The RBD and ACE2 structures in the optimized SARS-CoV RBD-ACE2 complex structure are shown as blue and green tubes, respectively. The location of noticeable subtle conformational difference is indicated by an arrow. C. Distinct interaction patterns in the SARS-CoV-2 and SARS-CoV RBD-ACE2 interfaces. Structures of RBD and ACE2 are shown as cartoon in pink and green colors, respectively. The side chains of the residues in both protein components, representing their unique interactions, are shown as sticks. Polar interactions (salt-bridge and hydrogen bond) are shown as blue dash line. Non-polar interactions (π-stack, π-anion, and hydrophobic interactions) are shown as orange dash line.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Sequencing

    Measurements of SARS-CoV-2 and SARS-CoV S1 binding to ACE2. A. Serial diluted recombinant S1 proteins of SARS-CoV-2, SARS-CoV and MERS-CoV were coated on 96 well plates, incubated with the recombinant Fc-tagged ACE2 (ACE2-Fc) for binding evaluation. B. Recombinant S1 proteins of SARS-CoV-2 and SARS-CoV were incubated with 293T-ACE2 cells and subjected to FACS evaluation for binding.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Measurements of SARS-CoV-2 and SARS-CoV S1 binding to ACE2. A. Serial diluted recombinant S1 proteins of SARS-CoV-2, SARS-CoV and MERS-CoV were coated on 96 well plates, incubated with the recombinant Fc-tagged ACE2 (ACE2-Fc) for binding evaluation. B. Recombinant S1 proteins of SARS-CoV-2 and SARS-CoV were incubated with 293T-ACE2 cells and subjected to FACS evaluation for binding.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Binding Assay, Recombinant, Incubation, FACS

    Conformational Effects of Trypsin Treatment of Spikes Follow the hACE2-Dependent Activation Pathway (A–C) The serine protease trypsin remodels conformational landscape of spike proteins toward down-stream conformations on the path of hACE2-dependent activation. (A and B) The FRET histogram (A) and TDP (B) of spike proteins on HIV-1 lentivirus particles in the presence of 50 μg/mL trypsin. (C) An experiment as in (A), for spikes in the presence of both 50 μg/mL trypsin and 200 μg/mL hACE2. (D) Three-dimensional presentations of FRET histograms of spike proteins on the virus in the presence and the absence of hACE2 and trypsin. FRET histograms represent mean ± SEM, determined from three randomly assigned populations of FRET traces. For evaluated state occupancies, see Table S1 . (E and F) Trypsin enhances SARS-CoV-2 spike-mediated hACE2-dependent virus-cell fusion. (E) Assay design to monitor virus-cell fusion using the HiBit and LgBiT split NanoLuc system ( Yamamoto et al., 2019 ). Vpr-HiBit was packaged into lentiviral particles carrying SARS-CoV-2 spike (LV_Spike). HEK293 target cells transiently expressing LgBiT tagged to PH domain of human phospholipase Cδ at the N terminus alone or together with hACE2. hACE2-dependent virus-cell fusion was determined by monitoring reconstituted NanoLuc activity in target cells 24 h after infection. (F) Normalized relative luciferase units (RLU; mean ± SD, two replicates with quadruplicates) measured 24 h post-infection to quantify virus-cell fusion in stated target cells after treating viruses with or with indicated amounts of trypsin for 15–20 min at 37°C. NanoLuc activities were normalized to luciferase activity detected in uninfected target cells. p values derived from unpaired t test; ∗∗∗∗ corresponds to p

    Journal: Cell Host & Microbe

    Article Title: Real-Time Conformational Dynamics of SARS-CoV-2 Spikes on Virus Particles

    doi: 10.1016/j.chom.2020.11.001

    Figure Lengend Snippet: Conformational Effects of Trypsin Treatment of Spikes Follow the hACE2-Dependent Activation Pathway (A–C) The serine protease trypsin remodels conformational landscape of spike proteins toward down-stream conformations on the path of hACE2-dependent activation. (A and B) The FRET histogram (A) and TDP (B) of spike proteins on HIV-1 lentivirus particles in the presence of 50 μg/mL trypsin. (C) An experiment as in (A), for spikes in the presence of both 50 μg/mL trypsin and 200 μg/mL hACE2. (D) Three-dimensional presentations of FRET histograms of spike proteins on the virus in the presence and the absence of hACE2 and trypsin. FRET histograms represent mean ± SEM, determined from three randomly assigned populations of FRET traces. For evaluated state occupancies, see Table S1 . (E and F) Trypsin enhances SARS-CoV-2 spike-mediated hACE2-dependent virus-cell fusion. (E) Assay design to monitor virus-cell fusion using the HiBit and LgBiT split NanoLuc system ( Yamamoto et al., 2019 ). Vpr-HiBit was packaged into lentiviral particles carrying SARS-CoV-2 spike (LV_Spike). HEK293 target cells transiently expressing LgBiT tagged to PH domain of human phospholipase Cδ at the N terminus alone or together with hACE2. hACE2-dependent virus-cell fusion was determined by monitoring reconstituted NanoLuc activity in target cells 24 h after infection. (F) Normalized relative luciferase units (RLU; mean ± SD, two replicates with quadruplicates) measured 24 h post-infection to quantify virus-cell fusion in stated target cells after treating viruses with or with indicated amounts of trypsin for 15–20 min at 37°C. NanoLuc activities were normalized to luciferase activity detected in uninfected target cells. p values derived from unpaired t test; ∗∗∗∗ corresponds to p

    Article Snippet: Construction of Full-Length Tagged SARS-CoV-2 Spike (S)A full-length wild-type pCMV3-SARS-CoV-2 Spike (S1+S2)-long (termed as pCMV-S, codon-optimized, Sino Biological, cat # VG40589-UT) plasmid was used as a template to generate tagged pCMV-S.

    Techniques: Activation Assay, Expressing, Activity Assay, Infection, Luciferase, Derivative Assay

    Antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2. a Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with COVID-19 human convalescent serum. Representative images of five experiments are presented. Scale bars: 50 µm. b Correlation analysis of neutralization of rVSV-∆G-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each serum sample ( n = 12), NT 50 values were determined for neutralization of rVSV-∆G-spike or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between the neutralization assays. Spearman’s correlation r and p values are indicated. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: A single dose of recombinant VSV-∆G-spike vaccine provides protection against SARS-CoV-2 challenge

    doi: 10.1038/s41467-020-20228-7

    Figure Lengend Snippet: Antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2. a Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with COVID-19 human convalescent serum. Representative images of five experiments are presented. Scale bars: 50 µm. b Correlation analysis of neutralization of rVSV-∆G-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each serum sample ( n = 12), NT 50 values were determined for neutralization of rVSV-∆G-spike or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between the neutralization assays. Spearman’s correlation r and p values are indicated. Source data are provided as a Source Data file.

    Article Snippet: Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full-length human codon-optimized S gene from pCMV3-SARS-CoV-2 S expression plasmid (Sino Biological, Cat# VG40588-UT) using the following primers: forward –atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

    Techniques: Infection, Staining, Neutralization

    Surface antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2: (A) Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with serum from COVID-19 human convalescent serum (right panel). (B) Correlation analysis of neutralization of rVSV-ΔG-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each sera (n=12), NT 50 values were determined for neutralization of rVSV-ΔG-spike, or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between th neutralization assays. R 2 =0.911.

    Journal: bioRxiv

    Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

    doi: 10.1101/2020.06.18.160655

    Figure Lengend Snippet: Surface antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2: (A) Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with serum from COVID-19 human convalescent serum (right panel). (B) Correlation analysis of neutralization of rVSV-ΔG-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each sera (n=12), NT 50 values were determined for neutralization of rVSV-ΔG-spike, or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between th neutralization assays. R 2 =0.911.

    Article Snippet: Plasmid construction The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

    Techniques: Infection, Staining, Neutralization