Dear ALL;
A few weeks ago, I posted my problem with MBP fusion protein.
Thank folks very much for the help. Here is my latest result.
1) Shortening the linker region would signifcantly reduce the cleavage of
target proteins from MBP;
2) Adding a C-terminal tag would minimize the degradation of target proteins
and facilitate the purification;
3) Low temperature induction(18 degree) would significantly reduce the soluble
aggregates;
However, all the optimization procedures resulted in a ~150KD-200KD oligomer
of MBP fusion protein after gel-filtration.
Only a very small portion of MBP fusion protein exsited as monomers even in the
expression condition with low temperature and 0.1mM IPTG.
Does anyone experience the oligomerization of MBP fusion protein?
Thanks again and have a nice weekend!
Jerry McCully
