3 replies to this topic

[aztecan princess]

Has anybody transtected SF9 (or other insect cell line) using lipofectamine in stead of cellfectine??

Has anybody expressed successfully a gen (GFP or other) directed by the CMV promoter in SF9 (or other insect cell line) using a virus or a plasmid as a vector?

[bob1]

I havn't tried insect cells, but CMV is a mammalian virus, I suspect that plasmids running off this promoter will not work in insect cells.

[aztecan princess]

I found only one paper where they infect with a virus expressing a GFP directed by CMV promoter, and they say fluorescence is weak but it’s there so CMVp is a little bit active. But they don’t show negative control, and I see a black background in the positive control (GFP directed by a insect cell promoter), and I see a very green background in the CMV-GFP transfection, what make me think may be what they see is just endogen fluorescence.

[mydove]

aztecan princess, on Feb 17 2009, 12:41 AM, said:

I found only one paper where they infect with a virus expressing a GFP directed by CMV promoter, and they say fluorescence is weak but it’s there so CMVp is a little bit active. But they don’t show negative control, and I see a black background in the positive control (GFP directed by a insect cell promoter), and I see a very green background in the CMV-GFP transfection, what make me think may be what they see is just endogen fluorescence.

Hope is not too late.
We used to tansfect S2 or insect cells with plasmid under Actin- or inducible-promoter(for example metallotheonein) control.