Baculovirus Expression System

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 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 1 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.

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