Baculovirus can not replicate in organisms other than insects, which is valued by researchers and been developed into BacMam expression system. The expression of the target gene in mammalian cells is mediated by the modified baculovirus. BacMam is an improved version of baculovirus expression system. The process from gene insertion to protein expression of BacMam is basically the same as that of BEVS, except that the ultimate host is mammalian cells rather than insect cells. Same as the BEVS, baculovirus expression in mammalian cells can be achieved in a variety of pathways, each pathway has its own advantages and disadvantages, and cannot replace each other.
Four types of BacMam expression services are available Creative BioMart. One of them is the BacMam + Bac to Bac Expression Approach, within which the BacMam vector is constructed through the procedure of Bac to Bac.
In BacMam system, the virus which contains target gene can be generated by the principle of Bac to Bac expression system. Tn7 mediated site-specific transposition was used to integrate the expression cassette from donor vector into the baculovirus backbone (bacmid) in E. coli DH10Bac. The recombinant bacmid is then extracted via the standard procedure and transformed into the insect cell to assemble and amplify. Finally, the amplified virus stock is transduced to mammalian cell lines.
In order to be identified by mammalian cells, a hCMV-IE1 promoter (or other mammalian promoter) is previously inserted into the donor vectors and it will be integrated to the baculovirus together with other elements as well as the gene of interest.
| Step | Service Description | Timeline | Deliverables | |
| 1 | BacMam Vector Construction |
|
1-2 weeks |
|
| 2 | Recombinant Baculovirus Production |
|
2-3 weeks |
|
| 3 | Propagate the mammalian host cell |
|
1.5 weeks |
|
The efficiency of BacMam transduction in a cell line was confirmed by expressing green fluorescent protein (GFP) using BacMam. It can be used to optimize the transduction conditions (such as medium, temperature, time, etc.)
Be used for membrane preparations or in functional assays
Be used for large-scale protein expression
The BacMam vector is constructed via Gateway recombination
Support multiple gene co-expression in host mammalian cell.
Creative BioMart has more than 10 years of experience in protein expression and is competent to satisfy various protein expression desire. We are expert in protein expression (intracellular or secretory), signal peptide selection, insect cell selection and culture. Choosing our BacMam Service, customers only need to provide the coding sequence of your target protein. We will provide customizes service from codon optimization to large-scale expression and purification. For more assistance, please contact us for help.