VSV-ΔG-GFP Plasmid Expression Vector
The plasmid pVSV-ΔG-GFP encodes the antigenomic-sense (or positive-sense) RNA of a replicaton-restricted recombinant vesicular stomatitis virus (rVSV) in which the glycoprotein (G) gene has been replaced with GFP. This plasmid is used together with plasmids encoding the VSV nucleocapsid (N), phosphoprotein (P), glycoprotein (G), and large polymerase subunit (L) to recovery VSV-G pseudotyped ΔG-GFP virus as described in [1]. The antigenomic RNA of ΔG-GFP is expressed from the bacteriophage T7 promoter in pBS, which has been further modified to contain the hepatitis delta ribozyme used to generate a precise 3' end of the VSV antigenomic RNA and a T7 terminator sequence cloned between the SacII and SacI restriction sites in pBS-SK+ [2, 3].
Recombinant vesicular stomatitis viru-ΔG (rVSV-ΔG) has been used to produce VSV pseudotypes containing the envelope glycoproteins of heterologous viruses including viruses that require high-level containment. Since the infectivity of rVSV-ΔG is restricted to a single round of replication, analyses of viral entry can be performed using just biosafety level 2 (BSL-2) containment.
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 Michael A. Whitt, Ph.D., University of Tennessee.
Part of The Investigator's Annexe program.
The plasmid pVSV-ΔG-GFP encodes the antigenomic-sense (or positive-sense) RNA of a replicaton-restricted recombinant vesicular stomatitis virus (rVSV) in which the glycoprotein (G) gene has been replaced with GFP. This plasmid is used together with plasmids encoding the VSV nucleocapsid (N), phosphoprotein (P), glycoprotein (G), and large polymerase subunit (L) to recovery VSV-G pseudotyped ΔG-GFP virus as described in [1]. The antigenomic RNA of ΔG-GFP is expressed from the bacteriophage T7 promoter in pBS, which has been further modified to contain the hepatitis delta ribozyme used to generate a precise 3' end of the VSV antigenomic RNA and a T7 terminator sequence cloned between the SacII and SacI restriction sites in pBS-SK+ [2, 3].
Recombinant vesicular stomatitis viru-ΔG (rVSV-ΔG) has been used to produce VSV pseudotypes containing the envelope glycoproteins of heterologous viruses including viruses that require high-level containment. Since the infectivity of rVSV-ΔG is restricted to a single round of replication, analyses of viral entry can be performed using just biosafety level 2 (BSL-2) containment.
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 Michael A. Whitt, Ph.D., University of Tennessee.
Part of The Investigator's Annexe program.
| Catalog Number | Product | DataSheet | Size | AVAILABILITY | Price | Qty |
|---|
| Product Type: | Plasmid |
| Alternative Name(s): | antigenomic-sense (or positive-sense) RNA of vesicular stomatitis virus (VSV) ΔG-GFP |
| Gene/insert name: | ΔG-GFP |
| Antibiotic Resistance: | Ampicillin or Kanamycin (please see vial label and packing slip) |
| Fusion Tag(s): | GFP |
| Concentration: | 100uL (100ng/uL) |
| Grow in E. coli at 37 C: | Yes |
| Cloning Site 5': | 5' VSV sequence joined directly to T7 promoter |
| Cloning Site 3': | 3' VSV sequence joined directly to HDV ribozyme |
| Insert Size: | 10,354 bp |
| Vector Backbone and Size: | pBS-SK-ΦT, 3105 bp |
| High or low copy: | High |
| Comments: | For suggested protocol, see: Whitt, MA, J. Virol. Methods, 2010. 169(2): p. 365-74. |
| Shipped: | Ambient temperature |
VSV recovery requires BHK-21 cells and T7 supplied by vaccinia virus infection.
- 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.
- Lawson, N.D., et al., Recombinant vesicular stomatitis viruses from DNA. Proc.Natl.Acad.Sci.(USA), 1995. 92(10): p. 4477-4481.
- Stillman, E.A., J.K. Rose, and M.A. Whitt, Replication and amplification of novel vesicular stomatitis virus minigenomes encoding viral structural proteins. J. Virol., 1995. 69: p. 2946-2953.
- Chiang CF, Flint M, Lin JS, Spiropoulou CF. Endocytic Pathways Used by Andes Virus to Enter Primary Human Lung Endothelial Cells. PLoS One. 2016 Oct 25;11(10):e0164768. View Article
- Acciani M, Alston JT, Zhao G, Reynolds H, Ali AM, Xu B, Brindley MA. Mutational Analysis of Lassa Virus Glycoprotein Highlights Regions Required for Alpha-Dystroglycan Utilization. J Virol. 2017 Aug 24;91(18). pii: e00574-17. View Article
- Willard KA, Alston JT, Acciani M, Brindley MA. Identification of Residues in Lassa Virus Glycoprotein Subunit 2 That Are Critical for Protein Function. Pathogens. 2018 Dec 26;8(1). View Article
- Baggen J, Persoons L, Vanstreels E, Jansen S, Van Looveren D, Boeckx B, Geudens V, De Man J, Jochmans D, Wauters J, Wauters E, Vanaudenaerde BM, Lambrechts D, Neyts J, Dallmeier K, Thibaut HJ, Jacquemyn M, Maes P, Daelemans D. Genome-wide CRISPR screening identifies TMEM106B as a proviral host factor for SARS-CoV-2. Nat Genet. 2021 Apr;53(4):435-444. View Article
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