Recombinant plasmid pBG68 produces Sindbis virus (SINV) replicons in cells that allows for fast and easy expression of your gene of interest.
The 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 structural proteins from the 5' subgenomic promoter were replaced with the Blasticidin-S-deaminase (BSD) gene for replicon selection. A unique XbaI restriction site 3' subgenomic promoter to allow genes of interest to be ligated into the replicon. Transfection of this construct into mammalian cells produces a self-replicating Sindbis virus genome that expresses the BSD protein and any genes of interest cloned into the unique XbaI site, that can be packaged into single-round infectious virus-like particles by co-transfection with packaging plasmid pBG256 that can be used to insert the replicon into poorly or non-transfectable cells (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:||9,280 bp|
|Vector Backbone and Size:||pcDNA3.1 Zeo+|
|High or low copy:||High|
|Storage:||Room Temperature (Dried), -20C (liquid)|
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