@phdthesis{Paul2004, author = {Paul, Annekatrin}, title = {M{\"o}glichkeiten und Grenzen der Forschung an embryonalen Stammzellen und des therapeutischen Klonens : eine Untersuchung der verfassungsrechtlichen und einfachgesetzlichen Aspekte}, series = {Verfassungsrecht in Forschung und Praxis}, volume = {16}, journal = {Verfassungsrecht in Forschung und Praxis}, publisher = {Kova?}, address = {Hamburg}, isbn = {3-8300-1395-7}, issn = {1616-9794}, pages = {181 S.}, year = {2004}, language = {de} } @article{HessSaffertLiebetonetal.2015, author = {Hess, Anne-Katrin and Saffert, Paul and Liebeton, Klaus and Ignatova, Zoya}, title = {Optimization of Translation Profiles Enhances Protein Expression and Solubility}, series = {PLoS one}, volume = {10}, journal = {PLoS one}, number = {5}, publisher = {PLoS}, address = {San Fransisco}, issn = {1932-6203}, doi = {10.1371/journal.pone.0127039}, pages = {14}, year = {2015}, abstract = {mRNA is translated with a non-uniform speed that actively coordinates co-translational folding of protein domains. Using structure-based homology we identified the structural domains in epoxide hydrolases (EHs) and introduced slow-translating codons to delineate the translation of single domains. These changes in translation speed dramatically improved the solubility of two EHs of metagenomic origin in Escherichia coli. Conversely, the importance of transient attenuation for the folding, and consequently solubility, of EH was evidenced with a member of the EH family from Agrobacterium radiobacter, which partitions in the soluble fraction when expressed in E. coli. Synonymous substitutions of codons shaping the slow-transiting regions to fast-translating codons render this protein insoluble. Furthermore, we show that low protein yield can be enhanced by decreasing the free folding energy of the initial 5'-coding region, which can disrupt mRNA secondary structure and enhance ribosomal loading. This study provides direct experimental evidence that mRNA is not a mere messenger for translation of codons into amino acids but bears an additional layer of information for folding, solubility and expression level of the encoded protein. Furthermore, it provides a general frame on how to modulate and fine-tune gene expression of a target protein.}, language = {en} } @misc{HessSaffertLiebetonetal.2015, author = {Hess, Anne-Katrin and Saffert, Paul and Liebeton, Klaus and Ignatova, Zoya}, title = {Optimization of translation profiles enhances protein expression and solubility}, series = {Postprints der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, journal = {Postprints der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, number = {518}, issn = {1866-8372}, doi = {10.25932/publishup-40957}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-409574}, pages = {14}, year = {2015}, abstract = {mRNA is translated with a non-uniform speed that actively coordinates co-translational folding of protein domains. Using structure-based homology we identified the structural domains in epoxide hydrolases (EHs) and introduced slow-translating codons to delineate the translation of single domains. These changes in translation speed dramatically improved the solubility of two EHs of metagenomic origin in Escherichia coli. Conversely, the importance of transient attenuation for the folding, and consequently solubility, of EH was evidenced with a member of the EH family from Agrobacterium radiobacter, which partitions in the soluble fraction when expressed in E. coli. Synonymous substitutions of codons shaping the slow-transiting regions to fast-translating codons render this protein insoluble. Furthermore, we show that low protein yield can be enhanced by decreasing the free folding energy of the initial 5'-coding region, which can disrupt mRNA secondary structure and enhance ribosomal loading. This study provides direct experimental evidence that mRNA is not a mere messenger for translation of codons into amino acids but bears an additional layer of information for folding, solubility and expression level of the encoded protein. Furthermore, it provides a general frame on how to modulate and fine-tune gene expression of a target protein.}, language = {en} }