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Center for Structural Biology

Expression Services

The facility currently utilizes E. coli , insect cell (baculovirus mediated), and mammalian cell expression systems. We have a library of expression vectors that users have access to for their projects. Typically, users provide a sequence-verified expression vector containing the gene(s) of interest. Alternatively, our staff can undertake expression vector construction if requested.

E. coli Expression

  • Small-scale test expressions in deep-well blocks can be conducted to determine optimal expression conditions
  • Large multi-liter shaking, suspension cultures of E. coli can be grown and processed at temperatures ranging from 18°C to 42°C. Max capacity is 18L at a time
  • 10L fermentation runs using our New Brunswick SIP Bioflow 415 instrument can be conducted when large quantities of cell mass are needed
  • The facility stocks a variety of E. coli strains to address specific issues like rare-codon usage, protein mis-folding, and expression in specialized media, for example, for the production of seleno-methione variants or 13C and 15N labeling

Baculovirus-Mediated Insect Cell Expression

  • Generation, titer determination, and storage of recombinant baculovirus
  • We grow and maintain both Sf9 and Hi-five insect cell lines
  • Small-scale test expressions in both suspension and adherent cultures can be conducted to determine optimal expression conditions
  • The facility has the capacity to carry out multi-liter, shaking suspension culture expression runs, max ~20L

Mammalian Cell Expression

  • We grow and maintain three main suspension cell lines cultured in minimal media: HEK 293-F (freestyle), CHO-S (freestyle), NS0.
  • We have a number of adherent cell lines that are used for specialized circumstances like limiting glycosylation or enabling DHFR stable cell selection.
  • We are able to generate stable, clonal cell lines expressing your protein of interest. We also have several strategies that we can use to optimize expression levels in these systems.
  • Small-scale test expressions in both suspension and adherent cultures via transient transfection can be conducted to determine optimal expression conditions.
  • We have the capacity to carry out large scale expression experiments (suspension and adherent), Max ~15L of suspension culture and ~3L of adherent culture in roller bottles.