title: RNA secondary structure and in vitro translation efficiency.
abstract: Cell-free protein synthesis is a promising technology featuring many advantages compared to in vivo expression techniques. However, most proteins are still synthesized in vivo due to relatively low protein yields commonly achieved in vitro, especially in the batch mode of reaction. In Escherichia coli S30 extract-based cell-free systems protein yields are supposed to be partially limited by a secondary structure formation of the mRNA. In this study we checked promising members of various classes of RNA chaperones and several different RNA helicases on their ability to enhance in vitro translation. The data clearly show that the addition of none of these factors provides a general solution to the problem. However, protein yields can be increased in presence of a microRNA hybridizing with the 5' untranslated region of mRNAs, possibly by inducing structural changes improving accessibility of the Shine Dalgarno sequence for the ribosomes.
authors.affiliation: Department of Biophysics and Physical Biochemistry, University of Regensburg, Postfach, D-93040 Regensburg, Federal Republic of Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Freischmidt
authors.forename: Michael
authors.initials: M
authors.lastname: Liss
authors.forename: Ralf
authors.initials: R
authors.lastname: Wagner
authors.forename: Hans Robert
authors.initials: HR
authors.lastname: Kalbitzer
authors.forename: Gudrun
authors.initials: G
authors.lastname: Horn
mesh_terms: D002474:Cell-Free System / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D035683:MicroRNAs / Q000378:metabolism; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D020365:RNA Helicases / Q000378:metabolism; D012329:RNA, Bacterial / Q000737:chemistry / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; D035683:MicroRNAs; D012329:RNA, Bacterial; D012333:RNA, Messenger; D020365:RNA Helicases