The plasmid pVSV-ΔG-PL 2.5 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 containing sequences for the following restriction enzymes (5'-MluI-KpnI-XhoI-XmaI-EagI-SphI-NheI-3'), which are all unique in the plasmid.
pVSV-ΔG-PL 2.5 can be used to insert a heterologous gene into the MCS for expression from the VSV genome. For example expression of a heterologous glycoprotein, a cytokine, or a reporter such as GFP or luciferase. Such vectors would be useful for vaccine studies, for use as a reporter virus, or in identification of host entry factors as described in .
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 . The antigenomic RNA of ΔG-PL 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.
Part of The Investigator's Annexe program.
|Gene/insert name:||ΔG-PL 2.5|
|Antibiotic Resistance:||Ampicillin or Kanamycin (please see vial label and packing slip)|
|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:||12,733 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.|
VSV recovery requires BHK-21 cells and T7 supplied by vaccinia virus infection.
If you publish research with this product, please let us know so we can cite your paper.