I want to express GFP protein in both sf9 and Aag2 insect cell line. We have cloned GFP protein in PIZ/V5-His insect expression vector. I got a very good GFP expression in sf9 cells (western blot). But I am not able to get GFP expression in Aag2 cells. Can anybody guide me in this regard or suggest more suitable insect expression vector.
Hi! Aag2 cells might not be compatible with the pIZ you are using since it contains a promoter suitable for expression in lepidopteran cell lines like Sf9, Sf21 etc. That may explain the successful expression in Sf9 and the lack of it in mosquito cells. It might be due to a fail in transfection or just incompatibility of the promoter, as I mentioned. I guess that you have to take into account the host-vector compatibility for your studies. If using Sf9 lines, I can recommend you vectors from the pOPIN suite which are also suitable for E. coli and mammalian cell expression. Also pFastBac works fine in my hands. Good luck!
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