Inclusion body formation of the recombinant protein expressed in pET28a in E.col
Posted 04 December 2012 - 08:44 AM
I am new to the forum and really looking forward to get help through this..........I have expressed a protein of nearly 60kd in pet 28a in BL21(E.coli) but I am getting it as inclusion body. Please suggest me some way to solubilise the protein. I have tried expression at 16 with low IPTG..
Posted 18 December 2012 - 03:33 AM
If you have not yet got ur protein in soluble fraction, here is a suggestion. You can try cloning the gene in pET 32a/b/c. it has thioredoxin tag which helps to get the protein in soluble fraction. I have similar proble with my expressed protein. I have tried several methods but none of them worked. Only cloning in pET32 can help.
Otherwise you can try supplying sorbitol and betaine in the expression media. Some time it might help.
Posted 09 January 2013 - 06:22 AM