Forskning ved Københavns Universitet - Københavns Universitet


A secretory system for bacterial production of high-profile protein targets

Publikation: Bidrag til tidsskriftTidsskriftartikel

  • Alexander Kotzsch
  • Erik Vernet
  • Martin Hammarström
  • Jens Berthelsen
  • Johan Weigelt
  • Susanne Gräslund
  • Michael Sundström
Escherichia coli represents a robust, inexpensive expression host for the production of recombinant proteins. However, one major limitation is that certain protein classes do not express well in a biologically relevant form using standard expression approaches in the cytoplasm of E. coli. To improve the usefulness of the E. coli expression platform we have investigated combinations of promoters and selected N-terminal fusion tags for the extracellular expression of human target proteins. A comparative study was conducted on 24 target proteins fused to outer membrane protein A (OmpA), outer membrane protein F (OmpF) and osmotically inducible protein Y (OsmY). Based on the results of this initial study, we carried out an extended expression screen employing the OsmY fusion and multiple constructs of a more diverse set of human proteins. Using this high-throughput compatible system, we clearly demonstrate that secreted biomedically relevant human proteins can be efficiently retrieved and purified from the growth medium.
TidsskriftProtein Science
Udgave nummer3
Sider (fra-til)597-609
Antal sider13
StatusUdgivet - 2011

ID: 40289591