Expression of protein in mammalian cells.

IndiraShukla Indira_Shukla at brown.edu
Thu May 25 15:33:41 EST 2000


Hi, I will appreciate any input in the following problem.

I am trying to express a 10kD protein which is fused to GFP protein
(27kD). I was able to have very good expresion only ONCE. I have not been
able to repeat the expression again. Even GFP alone does not express very
well. I am using AtT20 cells. We had a similar problem with GH3 cells. As
far as I know, I am doing everything the same as I did that one time
except the cell density, i did not count it that time, but I have tried
varying degree of confluency with no success. Adding Sodium butyrate does
not help. Can anyone with experience in expressing proteins in mammalian
cells give me soem pointers.

Thanks.






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