Insect Cell Lines
Vectors in Use
1. Baculovirus Expression System
Danieli's lecture notes.
Bac to Bac manual (PDF).
Expression System with Gateway Technology
tutorial (part of 72682 course material).
to Baculovirus Expression Vector systems (BEVS) and Insect Cell Culture
Lepidopteran cells: general, 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.
destination vectors for baculovirus expression system (PDF), list
- pAcGP67-C Baculovirus transfer vector
- MCS of PFastBac HC-1
- 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
Small scale purification of secreted His-tagged protein from conditioned medium
See also: Additional
|2. InsectSelect Stable
- pIB/V5 -His
- pIZT/V5 -His
- 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
MG1and KC1 cells.
vector search - type insectselect
|3. DES: Drosophila
Expression System For the stable expression and purification of heterologous
proteins in Schneider S2 cells.
The Drosophila Expression System.
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).
vector search - type pmt or pac.