Over Experssion of A Protein
Posted 03 December 2010 - 02:22 PM
My protein of interest is 296 a.a (MW 34.2kD). If I over express the full length of this protein it causes E. coli cell to die and little protein induction. However, if I over express either the C and N Terminus of this protein, we get nice induction. We tried with couple of E. coli strains to over express it and got the same results.
Thus, I would like to know what it is the reason behind this phenomenon.
E. coli: C41(DE3) also BL21(DE3)
Growth medium: 2xYT
OD: before induction 0.6
Any clue, please advice.
Posted 04 December 2010 - 04:50 AM
I have similar problems with my proteins...
You can try different temperatures. Are you doing the expression at 37ºC? You could try growing and expression at 28, or growing at 37 and expressing at 25 or even 16. I had success with the last combination...
Posted 06 December 2010 - 07:45 AM
talent does what it can
genius does what it must
i used to do what i got paid to do
Posted 06 December 2010 - 03:22 PM