Is your gene of interest smaller than 750bp ? Then I would synthesize the gene with IDT and optimize it for E.coli that's 139$ Jürgen
...................... Jürgen Bosch Johns Hopkins Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Phone: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-3655 http://lupo.jhsph.edu On Sep 12, 2013, at 10:23, "Elias Fernandez" <efern...@utk.edu<mailto:efern...@utk.edu>> wrote: 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
