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!!! :)
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.