BacMam System

Baculoviruses infects insects in the nature and is non-pathogenic to humans, but can transduce a broad range of mammalian and avian cells. BacMam system employs a baculovirus vector with the cytomegalovirus promoter to drive the expression of proteins in mammalian cells. Since insect viruses in mammalian cells are not multiplying, there is also no risk of infection to humans and the environment. BacMam system´s other advantages include

• high transduction rates; protein expression levels that can be adjusted by viral dose
• low cytotoxicity to host cells
• compatibility with a broad range of cells, including primary and stem cells
• ease and convenience of use

We have produced BacMam viruses mainly for different FRET biosensors, but also for YFP labeled Sigma 1 receptor and fluorescently labeled cell markers. These viruses have been used in different cell lines for example in B16F10 and HEK293 cells (Mazina et al. 2012), COS7 cells and JEG-3 (Mazina et al. 2015), KGN cells (Mazina et al. 2017).

Figure 1. The scheme of BacMam system example of using FRET biosensor. Firstly, the biosensor gene must be under a mammalian promoter, then the gene with the right promoter needs to be transformed into bacmid. The bacmid then gets transfected into Sf9 insect cells were the viruses reproduce. Then the that are intact are centrifuged and the supernatant, that contains viruses can be aliquoted. After that mammalian cells, that are infected with the virus and are producing biosensor can be used in an experiment in 96 well plate. In the final step, the data gathered for the experiment is analyzed.