BaculoGold System

Intracellular homologous is the presently preferred recombinant protein expression mode in BEVS. The recombinant process is completed by the co-transfecting of baculovirus DNA (linear or circular) and transfer vector into insect cell. However, the recombinant and infection process typically happen inside the insect cells which is invisible and cannot be monitored. To make the expression and infection process visualized, Creative BioMart provides a Bright Linearized BaculoGold system in which the baculovirus DNA framework is fused with a GFP tag. By fluorescence microscopy or flow cytometry, the recombinant viruses can be detected, allowing the study of kinetics of infection and protein expression.

How BaculoGold System Works

The rescue strategy is applied to this system. The BaculoGold DNA is engineered with a lethal deletion. Only when the BaculoGold DNA is co-transfected into the insect cell and homologous recombinant with the transfer plasmid which contain the segment of target gene, can the recombinant virus DNA survive in host insect cell. After that, driven by a promoter (which was simultaneously integrated to the virus DNA with target gene) insect cells began to express the target protein and GEP.

Figure 1. Expression Strategy of BaculoGold Approach

Our service for BaculoGold System BEVS

Here is the service process, which is very flexible. Customers can choose to start from or stop at any step they want. For more technical information or enquiry, please feel free to contact us.

Step		Service Description	Timeline	Deliverables
1	System Selection (Optional)	Select transfer vector Select host insect cell line (Sf9, Sf21, High Five)	——	——
2	Prepare Target Gene	Target gene sequence design and codon optimization Gene synthesis Subclone the interest sequence to the donor plasmid	< 1week	Enzymatic / Sequencing validation report.
3	Prepare Transfer Vector	Subclone the PCR product to the transfer vector Verify correct construct Extract recombinant vector	< 1week	Recombinant transfer vector Analyze report
4	Recombinant Baculovirus Production	Co-transfect insect cell with transfer vector and linear viral DNA Co-transfection of positive control Plaque assay and positive recombinant virus confirmation	1 weeks	Recombinant virus Recombinant rate report
5	Recombinant Virus Evaluation & Amplification	Prepare and assay of P1 virus Prepare P2 virus Virus titration	1.5 weeks	P1 and P2 virus stock Virus titration report
6	Optional Services	System optimization Secondary purification Tag removal Activity assay	1-2 weeks	Optimization solution Protein product Protein activity report
7	Large-Scale Expression & Purification (1-10L)	Culture amplification Western Blot Purification	1-3 weeks	Purified Protein QC report

Transfer Vector and Insect Cell Line

In order to optimize the expression approach, entry vectors and insect with unique advantage are available at Creative BioMart.

Transfer Vector

Transfer Vector	Use
pENTR/ TEV /D-TOPO	Directional TOPO cloning 5’ TEV sequence for N-terminal tag removal
pENTR/ D-TOPO	Directional TOPO cloning
pENTR/ SD /D-TOPO	Directional TOPO cloning Vector includes the Shine-Dalgarno sequence for E. coli expression-ready entry clone

Insect Cell Line

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

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 comprehensive 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.

Please note that all products/services provided are for research use only. Not intended for any clinical use.