Creative BioMart provides various vectors and cell lines as competent cells contents for the MultiBacMam expression system. Vector with different MCS, promoter and antibiotic resistance, E. coli strains and insect cell line with different elements and features are all listed below. Customers can check the corresponding descriptions and decide their intended ones.
Two types of acceptor vectors, pACEMam 1 and pACEMam 2, are currently available at Creative BioMart. In order to be recognized by mammalian cells, strong viral/mammalian promoters were added into the vector backbone previously. Currently, the promotors we utilized are cytomegalovirus (CMV) promoter and the hybrid CAG promoter.
The pACEMam 1 and 2 each harbor one MCS flanked by CMV or CAG promoter. The homing endonuclease site I–Ceul and BstXI site form a multiplication mode M, allowing integration of multiple gene cassettes. SV40 and β- A (β- action) are terminators. In addition, both vectors carry a ColE1 origin of replication for maintenance of high plasmid copy number. The whole expression sequence is encompassed with Tn7 which can induce transposition. There is also a LoxP site on the vector, facilitating the Cre-Lox recombination.
Three donor vectors are currently available at Creative BioMart. The over-all design of donor vectors is similar to that of acceptor vector, but there is no Tn7 site on donor vector, meaning that the donor vector needs to fusion with acceptor vector to complete the approach. In the case of donor vector, the HE site is consists of PI-Scel and BstXI. Different vector show resistance to different antibiotics, but all vectors contain a LoxP incomplete inverted repeat to create acceptor-donor or donor-donor fusions.
Again, in order to be recognized by mammalian cells, strong viral/mammalian promoters (CMV and CAG) were added into the vector backbone previously.
The DH10EMBacVSV E. coli strain is used in the MultiBacMam expression process. The bacterial cell contains a baculovirus genome backbone which is considered as the bacmid.
Note. The mCherry is only active in insect cells.
| Insect Cell | Advantage | Use |
| Sf9 (Spodoptera frugiperda) | Well-characterized host cell, good for plaquing | All general types of recombinant baculovirus production and protein expression |
| Sf21 (Spodoptera frugiperda) | High yield of intracellular protein | Intracellular recombinant baculovirus production and protein expression |
| High Five (Trichoplusia ni) | High yield of secreted protein, shorter culture period | Secreted protein expression |
The system can be used to transduce a wide range of mammalian cell lines including HEK293, HeLa, U2-OS, COS and CHO cells.
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 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.