The plasmid pVSV-FL+(2) encodes the antigenomic-sense (or positive-sense) RNA of vesicular stomatitis virus (VSV) 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+ [1, 2]. The reverse genetics method used to recover recombinant VSV (rVSV) from plasmids involves the transfection of vaccinia-T7 infected cells with 4 different plasmids; one that express the rVSV antigenome (pVSV-FL+(2)) and 3 other that express the VSV N, P, and L proteins, the cDNAs of which are also under control of T7 promoters.
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.
|Alternative Name(s):||antigenomic-sense (or positive-sense) RNA of vesicular stomatitis virus (VSV)|
|Antibiotic Resistance:||Ampicillin or Kanamycin (please see vial label and packing slip)|
|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:||11,161 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, spotted on filter paper|
VSV recovery requires BHK-21 cells and T7 supplied by vaccinia virus infection.
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