pOET6 BacMam Transfer Plasmid: Efficient Mammalian Expression via Baculovirus
The pOET6 BacMam Transfer Plasmid is a specialised baculovirus transfer vector designed to facilitate the expression of foreign genes in mammalian cells using the highly efficient BacMam system. Unlike traditional baculovirus vectors optimized for insect cell expression, pOET6 is engineered with a cytomegalovirus (CMV) immediate early gene promoter. This powerful promoter drives constitutive high-level gene expression in a wide range of mammalian cell lines, including CHO and HEK 293, allowing researchers to leverage the safety and scalability of baculovirus production for mammalian protein targets.
The plasmid is designed for seamless integration with the flashBAC™ Baculovirus Expression System or any other system that utilises homologous recombination at the polyhedrin locus in insect cells. Following cotransfection into insect cells (typically using TransIT®-Insect Reagent), the flanking AcMNPV and ORF1629 sequences facilitate recombination into the baculovirus genome. The resulting recombinant virus can be harvested and used to transduce mammalian cells in a titer-dependent manner. This system combines the ease of baculovirus generation with the post-translational modification capabilities of mammalian hosts.
Key Benefits:
- Mammalian Optimised: CMV Promoter. Features a potent CMV immediate early promoter to drive robust constitutive protein expression in mammalian cells.
- System Versatility: Broad Compatibility. Fully compatible with the flashBAC™ System and other baculovirus platforms relying on homologous recombination at the polyhedrin locus.
- Streamlined Cloning: Multiple Cloning Site. Includes a Multiple Cloning Site (MCS) with unique restriction sites for precise insertion of the gene of interest.
- Flexible Workflow: Dual Host System. Utilises insect cells for high-titer virus production and mammalian cells for final protein expression, offering a safe, replication-incompetent delivery method.
- Reliable Selection: Ampicillin Resistance. Contains an ampicillin resistance gene and Col E1 origin for effective selection and propagation in E. coli.
Technical Insights
Vector Map and Features
The pOET6 BacMam plasmid is constructed to ensure efficient replication and transfer. Key features include the bacterial Col E1 origin of replication (4169–4788 bp) and the Ampicillin resistance marker (4943–5803 bp) for maintenance in E. coli. The expression cassette is flanked by AcMNPV (329–1156 bp) and ORF1629 (2580–3921 bp) sequences to enable homologous recombination. The CMV promoter (1380–2188 bp) is positioned upstream of the MCS (2252–2303 bp), followed by an SV40 poly(A) signal (2313–2434 bp) to ensure proper transcript processing in mammalian cells.
Specifications
- Product Type: Transfer Plasmid (BacMam)
- Concentration: 500 ng/µl, sterile filtered in TE Buffer
- Volume: 20 µl
- Promoter: Cytomegalovirus (CMV) Immediate Early
- Selection Marker: Ampicillin (E. coli)
- Storage Conditions: Store at -20°C.
- Shelf Life: 1 year from the date of purchase when properly stored and handled.
- Ordering Information:
- E7-0250: 20 µl pOET6 BacMam Transfer Plasmid (MIR 6152)