The Tet-On systems

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wei
wei's picture
The Tet-On systems

I try to establish the eGFP experssion in the Tet-On system. I transfect the pRevTRE-eGFP vector into the BOSC cells, then I can get the virus which contain the pRevTRE-eGFP vector. Therefore, I can use the pRevTRE-eGFP vector to infect the 3T3 cells which had already contained the pRevTet-On vector. So, the inducible system will work by tetracycline. Now, I have a puzzle is while the BOSC cells are transfected by the pRevTRE-eGFP vector; I can see the green-fluorescence by pRevTRE-eGFP vector from the BOSC cells. But, that are not any inducisor (Tetracycline) and repressor at the BOSC, and I am sure the vector, cell, and every condition are right. Would somebody please solve the puzzle for me!? Thanks a lot!!! :)

samm
samm's picture
I'm not too very clear with

I'm not too very clear with your problem with the information you have provided, but it appears that you have leaky tet expression. You can test your serum for tetracycline/doxycycline, and use a fresh thaw for transfection. That can often help.