Marco Oliveira

Thank you CharonY for your response. I don't think that the protein is toxic, since other works have expressed homologues with no trouble. It isn't a membrane protein, and I'm not sure if it aggregates. It is a human protein. I think that the plasmid is stable as I add kanamicin during the induction. I may try to increase the antibiotics concentration a little bit.

Please, I need help. I bought a synthetic gene from Genscript which came already cloned in a expression vector, pET28a. I'm experienced in protein overexpression, I'm following all standard methods for protein overexpression. Other proteins are been overxpressed normally at my lab, however this particular one isn't. I double checked the sequence, the codons are optimized and everything looks fine. However, the protein does not express at all in BL21(DE3) background. Any clue or suggestion?