The protein interactome (or so-called multi-protein complexes) is reported to catalyze most of the key cellular functions structurally and functionally. Thus, the role of protein interaction has become the focus of contemporary biological research. However, some of the complexes are either present in low abundance or only exist temporarily in native cells. Multi-expression of heterologous proteins can be achieved by recombinant technology, but the construction process is labor-intensive, and provides no flexibility.
To address the difficulties, Creative BioMart provides the Multi Gene Expression Approach, specifically designed for overproduction of eukaryotic complexes via baculovirus expression vector system (BEVS).
Basic principle
The basic principle of heterologous protein multi-expression (Figure 1) is similar to that of conventional Bac to Bac. Several heterologous genes were inserted into the vector (acceptor and donor) and recombined with viral DNA by Tn7 transposon site in E. coli cells, and then the recombinant bacmid was transfected into insect cells to synthesize the target protein.
The multi gene expression approach consists of two vectors (donor and acceptor) while Bac to Bac only adopts donor vector. The complementary advantages of the two vectors can realize the rapid, flexible and efficient insertion of multiple genes.
Information of Multi Gene Expression Approach
Multi gene expression approach is an innovation method based on the conventional Bac to Bac expression. The general difference between two approaches are utilized vector and gene cassettes insertion strategy. The basic competent cell contents and recombinant strategy of Multi Gene Expression Approach are
| Multi Gene Approach | Bac to Bac Approach | |
| Vector | Donor vector + Acceptor vector | Donor vector |
| E. coli strains | pir+ cells, pir- cells, DH 10 E. coli | DH 10 E. coli |
| Insect cell | Sf9, Sf21, H5 | |
| Recombinant strategy | Homing endonuclease /BstXI cloning + Cre-Lox recombination + Tn7 transposition | Tn7 transposition |
| Heterologous cassettes insertion | 1 to n gene cassettes | 1 gene cassette |
Click here to find out the information of vectors, E. coli strains as well as insect cell lines corresponding to Creative BioMart’s multi gene expression service.
For the detail service guide of cassettes insertion and recombinant strategy selection, please click here
Here are the service contents available at Creative BioMart, please click the link below for more information.
| Step | Service Description | Timeline | Deliverables | |
| 1 | System Selection |
|
—— | —— |
| 2 | Generating vectors contains multigene expression cassettes |
|
2-3 weeks |
|
| 3 | Preparation of recombinant Bacmid |
|
1 week |
|
| 4 | Recombinant Baculovirus Production |
|
2 weeks |
|
| 5 | Small-Scale Expression & Purification |
|
1.5 weeks |
|
| 6 | Optional Services |
|
1-2 weeks |
|
| 7 | Large-Scale Expression & Purification (1-10L) (optional) |
|
1-3 weeks |
|
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 Multi Gene Expression 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.