Dear CCP4ers, We've been struggling with little (nearly none) expression of our protein, in both E coli and with in vitro transcription/translation methods. It appears that our mRNA has high 2' structure with a low dG (theoretically ~760kcal/mol). If this is indeed the source of our problem, are there any potential strategies to either disrupt the mRNA structure chemically (in E coli or in vitro) or with thermophile expression systems for expression at higher temperatures?
Regards, Elias
