BacMam Multi-Gene Expression Service

In mammalian cells, it is multi-protein complexes rather than individual protein molecules that conduct the key cellular functions. Therefore, a solution that can express the complex compound is required to facilitate the protein interactome research.

As the conventional multi-expression process can be labor-intensive and inflexible, Creative BioMart provides a multi-gene expression BacMam approach that supports flexible multiple exogenous fragments insertion and production of protein complexes in a wide range of mammalian cells.

Technology Description

BacMam multi-gene expression service can realize baculovirus-mediated multi-gene expression in mammalian cells.

(1). Several expression cassettes can be integrated into donor and acceptor vectors through HE and Cre/LoxP recombination (Figure 1).

(2). The recombinant vectors are then recombined with viral DNA backbone by Tn7 transposon site in E. coli cells to form bacmid.

(3). The recombinant bacmid is transfected into insect cells to synthesize the recombinant baculovirus. (4). The recombinant baculovirus can be administrated to the selected mammalian cell line and guide the form and assembling of the desired protein interactome. 

Note. In order to be recognized by mammalian cells, strong viral/mammalian promoters (currently CMV and the hybrid CAG promoters) need to be added into the vector backbone previously.

Technology Features

• The recombinant vectors can be constructed through either homing endonuclease (he) / BstXI multiplication or Cre-Lox recombination or standard fragment insertion via MCS. Mini-Tn7 elements support the site-specific transposition between bacmid and vector
• A CMV promoter from Cytomegalovirus is integrated into the BacMam vector to facilitate the gene expression in mammalian cells
• Chloramphenicol resistance gene, Ampicillin resistance gene and Gentamicin resistance gene for vector selection
• A vesicular stomatitis virus (VSV) peptide is integrated into the baculovirus genome previously to display the VSV peptide on the surface of the recombinant virus. Through this method, the mammalian cell transduction efficiency can be enhanced dramatically.
• A mCherry fluorescent protein is also inserted into the backbone genome of baculovirus to convenient the monitoring of virus amplification with the pink color. 

Our Services

Here are the service contents available at Creative BioMart. Please click the link below for more information. For customers who are interested in our BacMam expression service, please click the links below to learn more detail about our services. For any question or request, welcome to contact us or send us an online inquiry.

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