Higher Eukaryotic System
The baculovirus-insect cells system is widely used to produce recombinant proteins from a wide range of organisms. As opposed to E. coli expression system, it makes use of a higher eukaryotic cellular machinery and usually yields overexpressed proteins with proper folding and disulfide bonding. Cellular localization is usually identical to that of native protein and most post-translational modifications found in higher organisms have been shown to occur usually at the same locations. The latter cellular processings include signal peptide cleavage, N- and O-glycosilation, phosphorylation, acetylation, amidation, etc. The baculovirus system has proven to be one of the most successful aproach to produce milligram amounts of recombinant structurally- identical to the native counterpart and most often biologically active proteins.
Standard Service Description
All services include multiple steps from gene to purified protein as described in the table below.
| Description | Resulting Material | Average turnaround (*) |
| Gene Synthesis Stage | -Codon optimization -Gene synthesis -Cloning in standard pGB219 plasmid -Complete sequencing |
Cloned, synthesized and verified gene | 1-2 weeks |
| Subcloning in Baculovirus Transfer Vector Stage | -Plasmid prep -Subcloning Bac-donor vector -Clone selection -Complete sequencing |
Plasmid vector with cloned target gene | 2 weeks |
| Bacmid Generation Stage | -Plasmid prep -Recombinant bacmid selection -Recombinant bacmid DNA isolation -DNA Verification |
Bacmid DNA with cloned target gene | 2 weeks |
| Insect Cells Transfection Stage | -Multiple Sf9 transfection experiments -Virus titering -SDS-PAGE / Western blot analysis |
High titer recombinant baculovirus stock | 3 weeks |
| Pilot Expression Stage | -Expression pilot and optimization experiments -Determine yields and protein behavior |
10-50 µg of semi-purified protein | 1-2 weeks |
| Expression and Purification Stage | -Expression -Protein extraction -Protein purification -SDS-PAGE, WESTERN blot |
0.1 to 3 mg of purified protein Purity >95% |
2-3 weeks |







