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Protein Expression and Purification Core FacilityProtein Expression

Insect Cells / Baculovirus

Protein expression in the Baculovirus system

Expression of genes in insect cells is more time-consuming than in bacteria. Therefore, one should be prepared for the case that one construct fails or does not yield enough protein. Similar to E. coli, the success rate is higher if you have several constructs to test rather than trying to optimize a single construct. Additionally, in case of Baculovirus constructs, protein yields can vary with virus titer, expression time after infection, cell culture medium, etc.

We use insect cells for the expression of proteins that are difficult to express in E. coli. Our system of choice for generating the virus DNA containing the gene of interest is the Bac-to-BacTM system from Invitrogen.

For a general introduction and protocols for the Baculovirus expression system, we recommend reading the following user manuals from Invitrogen and Clontech, respectively:

Guide to Baculovirus Expression Vector Systems (BEVS) and Insect Cell Culture Techniques
BacPAK™ Baculovirus Expression System User Manual

For the generation of recombinant Baculovirus with your gene of interest, please download
Bac-to-Bac® Baculovirus Expression System

Cloning of a gene of interest into a suitable expression vector has to be done by the user and the correct sequence of the insert verified by sequencing before any expression experiments can be started. You can find suitable expression vectors in our vector list. We are happy to give advice ragarding the appropriate construct for your purpose.

Available insect cell lines:

  • Sf9
  • Sf21
  • High FiveT
  • ExpresSF+

Our service includes:

  • Transfection and selection for recombinant baculovirus
  • Amplification of high-titer virus stock
  • Small-scale time course expression test
  • Large-scale expression and purification of recombinant protein

Please find the charges for these steps under Services & Charges

Contact: pepcore@embl.de