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.

The Investigator's Annexe Part of The Investigator's Annexe program.

Catalog Number Product DataSheet Size AVAILABILITY Price Qty
EH1009
pVSV-ΔL-GFP Plasmid Expression Vector
10uL (1ug/uL) In stock
Regular Price:$685.00
On Sale:
EH1010
VSV-ΔL-GFP Plasmid Expression Vector System
w/ set of Helper Plasmids (VSV-N, VSV-P, VSV-L, VSV-G) In stock
Regular Price:$891.00
On Sale:
Specifications

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: 10uL (1ug/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

Data

VSV recovery requires BHK-21 cells and T7 supplied by vaccinia virus infection.

Provider
From the laboratory of Michael A. Whitt, Ph.D., University of Tennessee.
References
  1. 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.
  2. Lawson, N.D., et al., Recombinant vesicular stomatitis viruses from DNA. Proc.Natl.Acad.Sci.(USA), 1995. 92(10): p. 4477-4481.
  3. 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.

If you publish research with this product, please let us know so we can cite your paper.

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