title: Analyzing and enhancing mRNA translational efficiency in an Escherichia coli in vitro expression system.
abstract: The dependence of efficiency of translation initiation on mRNA sequence parameters was investigated in an Escherichia coli in vitro expression system. We designed a large-scale expression experiment focussing on the influence of sequence variations in the translated region (TR) of the mRNA without changing the 5'-untranslated region (5'-UTR). The level of translated protein from 756 expression constructs was measured and the influence of a large number of possible effector attributes was statistically analyzed. Base exchanges immediately adjacent to the start codon up to nucleotide (+)25 had a profound effect on translational efficiency. Correlation analysis revealed a significant dependence on base pair probability and G+C content on the expression level, indicating that mRNA secondary structure in this region hampers translation. Using our training data, we developed a methodology to predict and improve the translation efficiency of open reading frames (ORFs).
authors.affiliation: Biomax Informatics AG, Lochhamerstr. 11, Martinsried D-82152, Germany. Dieter.Voges@biomax.de
authors.forename: Dieter
authors.initials: D
authors.lastname: Voges
authors.forename: Manfred
authors.initials: M
authors.lastname: Watzele
authors.forename: Cordula
authors.initials: C
authors.lastname: Nemetz
authors.forename: Sabine
authors.initials: S
authors.lastname: Wizemann
authors.forename: Bernd
authors.initials: B
authors.lastname: Buchberger
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001482:Base Composition; D020029:Base Pairing; D001483:Base Sequence; D003062:Codon / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D008957:Models, Genetic; D009690:Nucleic Acid Conformation; D016366:Open Reading Frames / Q000235:genetics; D011336:Probability; D014176:Protein Biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012372:ROC Curve; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D003062:Codon; D012333:RNA, Messenger