pBG256 Recombinant Sindbis Virus Replicon Packaging Plasmid
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.
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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