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The Insect Baculovirus Expression Vector System (BEVS) is categorized as a eukaryotic expression system, distinguished by its high safety standards. With its expansive genome, it accommodates the insertion of large exogenous genes, offering a significant advantage in expressing proteins with substantial molecular weights.
Furthermore, it possesses the capability to accomplish full post-translational modifications and effectively express exogenous genes. The system comprises a transfer vector, baculovirus vector, and host cell components. Employing one or more potent baculovirus super-strong promoters, the system generates recombinant viruses by inserting the exogenous target gene into the promoter. This process achieves highly efficient expression of the exogenous gene as the recombinant viruses replicate within insect cells.
BEVS is commonly employed in the development of virus vaccines, such as those for influenza and HPV. Additionally, it is utilised in the production of cell signalling proteins, cytokines, and kinase development, among other applications.
Service Process
1. Plasmid construction
Process
- Codon optimization; gene synthesis
- The PCR product is ligated to the expression vectors
- Top 10 E.coli cells are transformed
- Obtain the correct recombinant plasmid
Features
Vector optimization - To enhance expression success and yield, we offer protein expression with C-terminal fusion tags alongside conventional N-terminal tags. This approach maintains protein bioactivity while ensuring high purity
Lead Time: 15-20 business days
2. Preparation of recombinant Bacmid and high titer virus
Process
- Transform DH10Bac cells to get recombinant Bacmid; PCR analysis; Isolate recombinant bacmid DNA
- Transfect insect cells with recombinant Bacmid DNA to produce baculovirus, and assess expression levels via SDS-PAGE. If needed, repeat the infection process
Features
Suspension transfection - The distinctive suspension transfection method significantly enhances protein expression levels and efficiently reduces the experimental timeframe
Lead Time: 15-20 business days
3. Scale up expression and purification
Process
- Infect insect cells with appropriate baculovirus
- The target protein is purified by affinity chromatography, ion exchange, hydrophobic and molecular sieves
Features
Following optimization, a substantial quantity of protein can be acquired by infecting host cells with low-passage virus
Lead Time: 5-10 business days
4. Optional additional services
Process
- Endotoxin removal, lyophilisation or filter-sterilisation
- Tag removal (Additional charge)
Features
Customers have the flexibility to select from a range of additional services tailored to their specific requirements
Lead Time: 5 business days
5. Quality control
Process
The purity, concentration and further tests are performed
Features
Detailed COA report is provided