I want to express a eukaryotic protein using E.coli. I assume that codon optimization may be helpful. I tried both GeneScript and ThermoFisher GeneArt. The two company generated different optimized DNA sequences for E.coli expression. Both claimed less GC content and better codons for E.coli to recognize for translation, while GeneScript also removed some possible restriction enzyme sites for protection purpose. How should I determine which optimization result is more reliable and better?
You should try it by attempting to express the protein using either sequence and comparing the results. There's no way to make additional progress by theorizing about it. If there was, then the two companies' sequences wouldn't disagree so widely. Genes are cheap, and testing two constructs is almost as little work as testing one. You're as ready as you're going to be. Go for it.
Thanks, John. I know that trying the two in the same time is going to be more efficient, but we are trying to save funding so maybe try one in a time. According to these two companies, synthesize our fragment (1000-1100nt) costs $300-400 (codon optimization included). Is this price reasonable?
Try geneart I have used it for over 100 genes while working in cro. It has worked all the time
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