The plasmid pBS-L-ΦT encodes the large subunit (L) of the vesicular stomatitis virus (VSV) polymerase protein expressed from the bacteriophage T7 promoter in pBS, which has been further modified to contain a T7 terminator sequence cloned between the SacII and SacI restriction sites in pBS-SK+ . The plasmid is used as part of the VSV reverse genetics system to recover recombinant VSV from plasmids [2, 3]. The recovery of rVSV involves the transfection of vaccinia-T7 infected cells with 4 (or 5) different plasmids; one that expresses the rVSV antigenome (e.g. pVSV-FL+(2) or pVSV-ΔG-GFP) and 3 (or 4) other plasmids that express the VSV N, P, and L proteins (or N, P, L and G for replication-restrictedΔG-VSV).
From the laboratory of Michael A. Whitt, Ph.D., University of Tennessee.
Part of The Investigator's Annexe program.
|Gene/insert name:||VSV large polymerase subunit (L)|
|Antibiotic Resistance:||Ampicillin or Kanamycin (please see vial label and packing slip)|
|Grow in E. coli at 37 C:||Yes|
|Cloning Site 5':||XhoI|
|Cloning Site 3':||XbaI|
|Insert Size:||6,446 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.|
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