iam student who just started work in biotechnology. now im facing big question, bout eucaryotic gene expression in procaryotic cells. there is a statement said, if i want to over express eucaryotic gene, so I have to remove its transmembrane domain. for your information, my gene is CWH41 gene which encoding membrane bound glycoprotein located in ER.
so my question is what is the affect of transmembrane domain during protein over expression in prokaryotic cells?
Check out the following thread if you haven't already done so. This topic has come up a couple of times. It can be difficult to express membrane proteins in e.coli (or other bacteria). For some reason, they are either poorly expressed and/or are toxic to the bacteria. In addition, the signal sequence may not be recognized, which means the protein is not corrected targeted to the membrane. I would assume once this happens the protein probably mis-folds and is either degraded or ends up in inclusion bodies. If you are just interested in the cytoplasmic portion of the protein, lets say you wanted to do pull-down assays to look at interaction partners, it would probably be easy to just express this portion of the protein.
As I mentioned, check out this thread and the following links: