We have created a modified toxin plasmid using a mammalian receptor domain. We are trying to express the modified toxin plasmid in cell culture, and isolate the protein. We need mammalian cell culture for glycosylation of the receptor domain. We are getting extremely low levels of expression. Any ideas?
Yes, you should worry about codon usage. If you do not, the most likely thing that you will get is extremely low expression levels (what you mention in your question)
Best
Jose A.
Codon optimization can increase yields by one hundred fold or more, not just a little bit. I never would have guessed it would make that much difference.
http://www.ncbi.nlm.nih.gov/pubmed/25768907
http://www.ncbi.nlm.nih.gov/pubmed/25926644
http://www.ncbi.nlm.nih.gov/pubmed/25665506
yes, codon optimization has to be done. it affects expression greatly
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