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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.
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|
-Cloning in standard pUC plasmid
|Cloned, synthesized and verified gene||1-2 weeks|
|Subcloning in Baculovirus Transfer Vector Stage|
-Subcloning Bac-donor vector
|Plasmid vector with cloned target gene||2 weeks|
|Bacmid Generation Stage|
-Recombinant bacmid selection
-Recombinant bacmid DNA isolation
|Bacmid DNA with cloned target gene||2 weeks|
|Insect Cells Transfection Stage|
|-Multiple Sf9 transfection experiments
-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|
-SDS-PAGE, WESTERN blot
| 0.1 to 3 mg of purified protein
(*) Average production time for average difficult project. Actual turn around time varies with individual project features.