Perguntas Frequentese de Clonagem e Expressão em LEXSY
Q12: A key problem in protein expression in E. coli is the insolubility of the recombinant proteins. Are the proteins expressed in LEXSY soluble and did you observe inclusion bodies?
A12: LEXSY has the advantage of a fully functional eukaryotic protein biosynthesis, folding and modification machinery. A eukaryotic - especially mammalian - target protein is expected to fold more natively in Leishmania than in E. coli. All proteins we tested so far were soluble and proteins insoluble in E. coli were expressed in soluble form in LEXSY (for examples see p.18 of the LEXSY brochure 2011 and the chapter “Selected Applications” of the LEXSY section of the Jena Bioscience website). In addition to folding, LEXSY performs mammalian-type posttranslational modifications as N-glycosylation which may be essential for activity, function and stability of the target protein.