VSV-ΔG-P/M-MCS2-2.6 Plasmid Expression Vector

The plasmid pVSV-ΔG-P/M-MCS2-2.6 encodes the antigenomic-sense (or positive-sense) RNA of a replication-restricted recombinant vesicular stomatitis virus (rVSV) in which the glycoprotein (G) gene has been deleted and replaced with a multiple cloning site (MCS-1) containing the following restriction enzyme sites (5'-MluI-KpnI-NheI-3') which are unique in the plasmid. It also contains an additional transcription start-stop sequence between the P and M genes, which has a second multiple cloning site (MCS-2) that has sites for the enzymes (5'-AscI-NotI-AvrII-3'). These are also unique in the plasmid.

pVSV-ΔG-P/M-MCS2-2.6 can be used to insert two heterologous genes for expression from the VSV genome. For example expression of a heterologous glycoprotein from MCS-1 and expression of a cytokine or a reporter from MCS-2. When recovered, these viruses would be useful for vaccine studies, use as reporter viruses in high-throughput antiviral drug screening, or in identification of host entry factors as described in [1].

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-virus as described in [1]. The antigenomic RNA of ΔG- P/M VSV is expressed from the bacteriophage T7 promoter in pBluescript, 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].

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.

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Catalog Number Product DataSheet Size AVAILABILITY Price Qty
EH1032
VSV-ΔG-P/M-MCS2-2.6 Plasmid Expression Vector
10uL (1ug/uL) In stock
Regular Price:$702.00
On Sale:
EH1033
VSV-ΔG-P/M-MCS2-2.6 Plasmid Expression Vector System
w/ set of Helper Plasmids (VSV-N, VSV-P, VSV-L, VSV-G) In stock
Regular Price:$913.00
On Sale:
Specifications

Product Type: Plasmid
Gene/insert name: ΔG-P/M-MCS2-2.6
Antibiotic Resistance: Ampicillin or Kanamycin (please see vial label and packing slip)
Fusion Tag(s): None
Concentration: 10uL (1ug/uL)
Amount: 10 ul aliquot of plasmid in shipping vial
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: 12,792 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

pVSV-DeltaG-GFP-2.6

Provider
From the laboratory of Michael A. Whitt, Ph.D., University of Tennessee.
Comments

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

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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