Recombinant plasmid pBG218 expresses Sindbis virus (SINV) expressing GFP during viral replication for easy visualization.
The entire Sindbis virus genome was inserted into the pcDNA3.1 Zeo+ expression vector such that the transcription start site of the CMV promoter overlaps with the first viral nucleotide of the viral genome. A Hepatitis D virus (HDV) ribozyme was added to the 3' end of the viral genomic 35bp polyA sequence. The gene for GFP was behind the 3' subgenomic promoter to allow expression of GFP protein during viral infection. Transfection of this construct into mammalian cells produces a self-replicating Sindbis virus genome that expresses GFP only when the virus genome is replicated in vitro or in vivo.
From the laboratory of Brian J. Geiss, PhD, Colorado State University.
|Gene/insert name:||Sindbis virus replicon|
|Organism:||Sindbis Virus Strain TE3'2J (AR339)|
|Grow in E. coli at 37 C:||DH5-alpha or XL10-Gold E. coli|
|Cloning Site 5':||MluI|
|Cloning Site 3':||XhoI|
|Insert Size:||13,363 bp|
|Vector Backbone and Size:||pcDNA3.1 Zeo+|
|High or low copy:||High|
|Storage:||Room Temperature (Dried), -20C (liquid)|
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