4 replies to this topic

[dc11]

Hi all,

If a specific protein is strongly expressed in a cell type, can I still overexpress this protein in the cell using a plasmid transient transfection? Thanks a lot in advance for your input.

dc11

[bob1]

You should be able to as the plasmids typically run off a viral, constitutively expressed promoter.

[Bomber]

You can, but it might end up in having no apparant effect on the cell.
In case you haven't done it yet, you might want to repress the expression and then check for phenotypes.

[warsel]

I don't know of any case where that isn't possible. Even if your protein is already highly expressed, a constitutive powerful promoter (like CMV for instance) should be able to boost expression.

If you will see a phenotypic difference though is a different matter (and more so if your phenotypic difference is due to overexpression of ANY protein* or due to specific effects of the protein you chose to overexpress)

*like accumulation of misfolded proteins, putting stress on the protein synthesis machinery, aggregation etc.

Edited by warsel, 06 June 2009 - 02:21 AM.

[giota22]

Hello, I'm new to this forum. Could you analyze to me what protein overexpression is, where it is used and everything relevant to this subject. Thank you!