VSV-ΔL-GFP Plasmid Expression Vector
The plasmid pVSV-ΔL-GFP encodes the antigenomic-sense (or positive-sense) RNA of a non-replicating recombinant vesicular stomatitis virus (rVSV) minigenome in which the polymerase (L) gene has been replaced with GFP. This plasmid is used as a transfection control in VSV recovery experiments as described in [1]. The antigenomic RNA of ΔL-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]. The reverse genetics method used to recover recombinant VSV (rVSV) from plasmids involves the transfection of vaccinia-T7 infected cells with 4 to 5 different plasmids; one that expresses the rVSV antigenome and 3 (or 4) other plasmids that express the VSV N, P, and L proteins (or N, P, L and G proteins if recovering a replication-restrictd ΔG-VSV).
Recombinant vesicular stomatitis virus-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-ΔL-GFP encodes the antigenomic-sense (or positive-sense) RNA of a non-replicating recombinant vesicular stomatitis virus (rVSV) minigenome in which the polymerase (L) gene has been replaced with GFP. This plasmid is used as a transfection control in VSV recovery experiments as described in [1]. The antigenomic RNA of ΔL-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]. The reverse genetics method used to recover recombinant VSV (rVSV) from plasmids involves the transfection of vaccinia-T7 infected cells with 4 to 5 different plasmids; one that expresses the rVSV antigenome and 3 (or 4) other plasmids that express the VSV N, P, and L proteins (or N, P, L and G proteins if recovering a replication-restrictd ΔG-VSV).
Recombinant vesicular stomatitis virus-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 |
| Gene/insert name: | VSV-ΔL-GFP |
| Antibiotic Resistance: | Ampicillin or Kanamycin (please see vial label and packing slip) |
| Fusion Tag(s): | none |
| 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: | 5,540 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.
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