Presently, the preferred recombinant protein expression mode is intracellular homologous recombination which is conducted by co-transfecting baculovirus DNA and transfer vector into insect cell. However, that plaque purification after recombinant vector transferred into insect cell can be time-consuming.
Creative BioMart provides a faster baculovirus-based expression system, which eliminates the labor-intensive plaque screening and purification step. By removing some essential portion from framework of viral DNA, the recombinant rate can be improved to nearly 100% thus avoid the screening and purification. Three kinds of BacMagic DNA (virus DNA) frameworks and transfer vectors are available at Creative BioMart. By the different matching between combinations of DNA and vectors, all the demands including high-level expression, secretory protein expression, C/N-terminal dual tag expression, fusion protein expression, multisubunit protein expression, and baculovirus surface display can be satisfied.
The rescue strategy is applied here to eliminate the non-recombinant background (Figure 1). The essential ORF 1629 fragment is partially removed while the polyhedrin coding region is replaced by bacterial artificial chromosome (BAC). This strategy allows circular DNA multiplying in bacteria but prevent non recombinant virus from replicating in insect cells. The homologous recombination between the virus DNA framework and transfer vector can restore the missing part of ORF as well as replace the BAC. Therefore, all viable baculovirus are recombinant-positive virus. Of course, in order to verify the effect of recombination, researchers can require a plaque assay after co-transfection. Creative BioMart will always be willing to try our best to provide customized service for you.
Figure 1. Basic Expression Process of BacMagic Expression Approach
Here is the general service process. For more technical information or enquiry, please feel free to contact us.
| Step | Service Description | Timeline | Deliverables | |
| 1 | System Selection （Optional） |
|
—— | —— |
| 2 | Prepare Target Gene |
|
< 1week |
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| 3 | Recombinant Virus Generation |
|
5 days |
|
| 4 | Recombinant Protein |
|
3 days |
|
Generally, the BacMagic system is referred to
BacMagic DNA
Here are three BacMagic DNA options. In a unified way, part of ORF1629 and polyhedrin coding regions were deleted and replaced by BAC, respectively. Other features and use have all been listed below. Customers can either choose the one that fit your protein best or contact us for help.
| Transfer Vector | Feature | Use |
| BacMagic-1 DNA |
|
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| BacMagic-2 DNA |
|
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| BacMagic-2 DNA |
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Transfer Vector
| Transfer Vector | Feature |
| pBAC |
|
| pIEx/Bac |
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| pTtiEX |
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Creative BioMart offers vector with various function. We have N/C-terminal fusion tag vector, tag-free vector and LIC-prepared vectors under each category of transfer vector. Please click here to see our vector gallery and get the guidance of vector selection.
Insect Cell Line
Sf9, Sf21, S2, and High Five cells are all suitable for the InsectDirect system while the best result can always be obtained with Sf9, Sf21 cell line. Please click here to learn more about the feature and use of the host insect cell lines.
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 comprehensive 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.