Para la investigación
The baculovirus-insect cells system is widely used to produce recombinant proteins from a wide range of organisms. 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.
Service Description
All services include multiple steps from gene to purified protein as described in the table below.
Leadtime is approximately 10-14 weeks.
Description | Resulting Material | Average turnaround (*) |
Gene Synthesis Stage | ||
-Codon optimization -Gene synthesis -Cloning in standard pUC 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 |
(*) Average production time for average difficult project. Actual turn around time varies with individual project features.