I used the pF25K ICE T7 Flexi Vector from Promega with the T7 in vitro transcription/translation kit for small scale protein expression. Now I need to scale up my expression and my PI suggested transfecting pF25K into insect cells instead of cloning coding region into baculovirus to save some time. Has anyone ever done this with a plasmid that was originally designed for in vitro translation?
I have used the cell lines for baculovirus expression, and the vector you plan to use will not likely express protein in them. The only reason it works in the insect cell lysate system is because it has added T7 polymerase, a bacteriophage polymerase that works with the T7 promoter in the vector. You need a polyhedrin promoter to get high expression form the cells, the polyA site will not be enough. If you have an insect line already expressing T7 polymerase, it would work, but not the standard Sf9 or Sf21 cells.
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