The plasmid pBS-G-ΦT encodes the vesicular stomatitis virus (VSV) glycoprotein (G) from the Indiana serotype 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 replication-restricted ΔG-VSV from plasmids . The recovery of ΔG-VSV involves the transfection of cells that have been infected with recombinant vaccinia expressing bacteriophage T7 RNA polymerase (vvT7) with 5 different plasmids; one which expresses the ΔG-VSV antigenomic-sense (or positive-sense) RNA and 4 other plasmids that encode individually the VSV N, P, G and L proteins, all of which are under control of T7 promoters.
From the laboratory of Michael A. Whitt, Ph.D., University of Tennessee.
Part of The Investigator's Annexe program.
|Gene/insert name:||VSV-Indiana glycoprotein (G)|
|Antibiotic Resistance:||Ampicillin or Kanamycin (please see vial label and packing slip)|
|Grow in E. coli at 37 C:||Yes|
|Cloning Site 5':||ApaI|
|Cloning Site 3':||BamHI|
|Insert Size:||1,700 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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