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Protein Expression 
in Insect Cells

baculovirus infected sf21 cells
Baculovirus-infected  SF21 cells

Expression System
Insect Cell Lines
Vectors in Use
Related Procedures

1. Baculovirus Expression System

  • Dr. Danieli's lecture notes.
  • Invitrogen's Bac to Bac manual (PDF).
  • Baculovirus Expression System with Gateway Technology
  • The Baculovirus tutorial (part of 72682 course material).
  • Guide to Baculovirus Expression Vector systems (BEVS) and Insect Cell Culture Techniques
  • DH10Bac 
  • Lepidopteran cells: general, SF9 (PDF), SF21, High-5, Mimic-SF9 (PDF),
  • Baculovirus can infect Drosophila cells, however the virus will not replicate in the infected cells and will not be able to infect subsequent cells. No high-titer stocks can be generated from Drosophila cells.
  • Gateway destination vectors for baculovirus expression system (PDF), list
  • General vectors:
    1. FlashBac
    2. pAcGP67-C Baculovirus transfer vector
  • Bac-To-Bac vectors:
    1. MCS of PFastBac HC-1
    2. pFastBac1 - map, MCS
  • Cryopreservation of SF9/H5 cells
  • Insect cells cryopreservation (S9, Sf21, S2, Tni)
  • Rapid thaw of SF9 and H5 cells
  • Transfection of Bacmid DNA into SF9 cells using ESCORT reagent
  • Transfection of FlashBac DNA & transfect vector into SF9 cells using ESCORT reagent.
  • Viral plaque assay
  • Viral plaque assay, abridged
  • Amplification of purified plaques.
  • Boosting protein expression in High5 & SF9 cells
  • Selenomethionine incorporation
  • Small scale purification of secreted His-tagged protein from conditioned medium

  • See also: Additional resources
    2. InsectSelect Stable Expression System
  • InsectSelect - pMIB/V5-His 
  • InsectSelect - pIB/V5 -His
  • InsectSelect - pIZT/V5 -His 
  • InsectSelect -  pIZ/V5 - HIS
  • The InsectSelect System uses the OpIE2 promoter from the Orgyia pseudotsugata multicapsid nucleopolyhedrosis virus for high-level expression. This promoter is active in many common insect cell types including SF9, SF21 , High Five™, Mimic-SF9,S2, MG1and KC1  cells.
  • Invitrogen vector search - type insectselect
  • pMIB/V5-His (map, manual)

    See: Additional resources
    3. DES:  Drosophila Stable Expression System Drosophila Expression System For the stable expression and purification of heterologous proteins in Schneider S2  cells.

    The Drosophila Expression System.
  • S2 Schneider cells
  • D.Mel-2 cells (protocols) These cells were  isolated from late-stage Drosphila melanogaster embryos and adapted to suspension growth in serum free medium.
  • The drosophila MT promoter used in this system does not work well in lepidopteran cells (Sf9, sF21, and High Five). 
  • Invitrogen vector search - type pmt or pac. 
  • pMT/BiP/V5-His (map, manual)
  • See: Additional resources

    This site is maintained by Dr. Nurit Doron . Your comments are most welcome.

    entries since October 2003

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