Recombinant packaging plasmid pBG256 lacking the virla nsP1-nsP4 replication genes and can be used to combine plasmids with pBG60, pBG68 and pBG78.
The Sindbis virus genome lacking the viral nsP1-nsP4 replication genes and possessing the viral structural protein genes under the control of the native viral subgenomic promoter 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 35 base polyA sequence. Transfection this construct into mammalian cells produces a Sindbis virus genome that cannot express the viral structural proteins unless the viral nsP1-nsP4 replication proteins are provided in trans by a co-transfected replicon plasmid (pBG60, pBG68, pBG78). This construct was found to be superior to the pBG44 packaging construct described in Geiss et al., 2007 in production of virus-like particles. Equimolar amounts of replicon and packaging plasmids should be transfected into cells for optimal results.
From the laboratory of Brian J. Geiss, PhD, Colorado State University.
Product Type: | Plasmid |
Gene/insert name: | Sindbis virus replicon packaging plasmid |
Organism: | Sindbis Virus Strain TE3'2J (AR339) |
Antibiotic Resistance: | Amp |
Grow in E. coli at 37 C: | DH5-alpha or XL10-Gold E-coli at 37C |
Cloning Site 5': | MluI |
Cloning Site 3': | XhoI |
Insert Size: | 5,476 bp |
Vector Backbone and Size: | pcDNA3.1 Zeo+ |
High or low copy: | High |
Storage: | Room Temperature (Dried), -20C (liquid) |
Shipped: | Room Temperature |
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