Hi all !!
I have big trouble expression my protein in E coli.
my protein is a DNA modifying enzyme, and I believe that is the problem : its toxicity.
The DNA seq is correct. I also add one extra stop codon (TAA) after the original one which is an TAG.
I used different e coli lines, also the rosetta2 .
I used induction at 37 C different times and RT up to over night.
I also tried to incubate the bacteria in protease inhibitors to avoid protein degradation.
any idea ?
thank you in advance for ur help