The Baculovirus expression vector system (BEVS) has been developed since 1983 (Smith & Summer, 1983; Maeda et al 1984), due to its high expression ability, safety, easy operation and other characteristics, it has become a powerful and popular tool for the research and production of various prokaryotic and eukaryotic proteins, and has successfully expressed a large number of exogenous genes, almost covering most of the genes that can be developed and utilized in all species.

1. Plan design - analyze target gene sequence and make experimental plan according to customer requirements and existing literature reports. Our company can provide customers with: pFASTBAC Ⅰ intracellular expression vector and our self-modified pFAST-GP67 and pFAST-HBM secretion expression vector for free;

2. Construction of recombinant donor plasmid "pFastBAC Ⅰ-target" (target represents target gene) was constructed after 1-2 weeks with appropriate vectors and labels (eg:His,GST, FC, SUMO).

3.The recombinant plasmid was transformed into DH10BAC competent cells one week. The positive clones were screened by blue and white spots and identified repeatedly by PCR. Finally, the recombinant plasmid was extracted by alkali lysis.

4. cells were transfected into SF21 cells after 1 week of recovery and subcultured in suspension. The recombinant rods were transfected into SF21 cells using chemical transfection reagent to prepare P1 generation of recombinant baculovirus.

5. Expression of recombinant protein One week, P2 virus was prepared by P1 baculovirus, and P2 virus was used to infect SF21 cells in large-scale culture to express recombinant protein.

6. Separation and purification of recombinant proteins and QC detection for 1 week. Use appropriate chromatographic methods (EG: affinity, ion, size exclusion, etc.) to isolate and purify the recombinant proteins, and ultrafiltration solution is changed to the buffer solution specified by the customer (PBS or Tri-HCl is selected as the final storage buffer if there is no special requirement). QC tests included: SDS-PAGE electrophoresis to detect the purity of the recombinant protein, BCA quantitative concentration of the recombinant protein, and Western-blot to detect the immunogenicity of the recombinant protein.