pVSV-Δ-G-SARS-CoV-2-S_nLucP Plasmid
The plasmid pVSVΔG-SARS-CoV-2Spike_Met1-D614Gdelta21-NLucP encodes the antisense RNA of a replication competent chimeric vesicular stomatitis virus (rVSV) in which the glycoprotein (G) has been replaced with the Spike protein of SARS-CoV-2. The Spike protein contains the D614G amino acid substitution and the last 21 residues of the cytoplasmic tail have been removed to enhance Spike protein incorporation onto particles. The genome also includes a nano-luciferasePEST reporter gene for easy monitoring of viral replication. This plasmid can be transfected into cells along with Helper Plasmids encoding the VSV nucleocapsid (N), phosphoprotein (P), glycoprotein (G), and large polymerase subunit (L) to recover rVSVΔG-SARS-CoV-2Spike_Met1-D614Gdelta21-NLucP virus as described in [1-3].
Recombinant vesicular stomatitis virus (rVSVΔG) has been used to produce VSV chimeric viruses containing the envelope glycoproteins of heterologous viruses including viruses that require high-level containment. These particles are capable of multi-cycle replication.
It is the responsibility of the principal investigator to seek Institutional Biosafety Safety Committee approval for recombinant DNA, transgenic animal or infectious agent use within their laboratory spaces and maintain an Institutional Biosafety Safety Committee approval during the time period these materials are used.
From the laboratory of Melinda A. Brindley, PhD, University of Georgia.
The plasmid pVSVΔG-SARS-CoV-2Spike_Met1-D614Gdelta21-NLucP encodes the antisense RNA of a replication competent chimeric vesicular stomatitis virus (rVSV) in which the glycoprotein (G) has been replaced with the Spike protein of SARS-CoV-2. The Spike protein contains the D614G amino acid substitution and the last 21 residues of the cytoplasmic tail have been removed to enhance Spike protein incorporation onto particles. The genome also includes a nano-luciferasePEST reporter gene for easy monitoring of viral replication. This plasmid can be transfected into cells along with Helper Plasmids encoding the VSV nucleocapsid (N), phosphoprotein (P), glycoprotein (G), and large polymerase subunit (L) to recover rVSVΔG-SARS-CoV-2Spike_Met1-D614Gdelta21-NLucP virus as described in [1-3].
Recombinant vesicular stomatitis virus (rVSVΔG) has been used to produce VSV chimeric viruses containing the envelope glycoproteins of heterologous viruses including viruses that require high-level containment. These particles are capable of multi-cycle replication.
It is the responsibility of the principal investigator to seek Institutional Biosafety Safety Committee approval for recombinant DNA, transgenic animal or infectious agent use within their laboratory spaces and maintain an Institutional Biosafety Safety Committee approval during the time period these materials are used.
From the laboratory of Melinda A. Brindley, PhD, University of Georgia.
| Product Type: | Plasmid |
| Gene/insert name: | VSVΔG-SARS-CoV-2Spike_Met1-D614Gdelta21-NLucP |
| Fusion Tag(s): | NLucP |
| Grow in E. coli at 37 C: | Yes |
| Selectable markers: | AmpR |
| Insert Size: | 14109bp |
| Vector Backbone and Size: | pBS-SK-ΦT, 3105bp |
| High or low copy: | High |
| Storage: | -20C |
| Shipped: | Room temperature |
- Whitt, M.A., Generation of VSV pseudotypes using recombinant DeltaG-VSV for studies on virus entry, identification of entry inhibitors, and immune responses to vaccines. J. Virol. Methods, 2010. 169(2): p. 365-74
- Havranek KE, Jimenez AR, Acciani MD, Lay Mendoza MF, Reyes Ballista JM, Diaz DA, Brindley MA. SARS-CoV-2 Spike Alternations Enhance Pseudoparticle Titers and Replication-Competent VSV-SARS-CoV-2 Virus. Viruses. 2020 Dec 18; 12(12). PMID: 33353101
- Lay Mendoza MF, Acciani MD, Levit CN, Santa Maria C, Brindley MA. Monitoring viral entry in real-time using a luciferase recombinant vesicular stomatitis virus producing SARS-CoV-2, EBOV, LASV, CHIKV, and VSV glycoproteins. Viruses. 2020 Dec 17; 12(12). PMID: 33348746
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