Author: Cobucci-Ponzano, Beatrice; Conte, Fiorella; Benelli, Dario; Londei, Paola; Flagiello, Angela; Monti, Maria; Pucci, Piero; Rossi, Mosè; Moracci, Marco
Title: The gene of an archaeal a-l-fucosidase is expressed by translational frameshifting Document date: 2006_8_18
ID: 69gftii4_24
Snippet: In vitro translation assays were performed essentially as described by Condò et al. (28) . The samples (25 ml final volume) contained 5 ml of S.solfataricus cell extract, 10 mM KCl, 20 mM Tris-HCl, pH 7.0, 20 mM Mg acetate, 3 mM ATP, 1 mM GTP, 5 mg of bulk S.solfataricus tRNA, 2 ml of [ 35 S]methionine (1200 Ci/mmol at 10 mCi/ml) and $10 pmol of each mRNA. The mixtures were incubated at 70 C for 45 min. After this time, the synthesized proteins .....
Document: In vitro translation assays were performed essentially as described by Condò et al. (28) . The samples (25 ml final volume) contained 5 ml of S.solfataricus cell extract, 10 mM KCl, 20 mM Tris-HCl, pH 7.0, 20 mM Mg acetate, 3 mM ATP, 1 mM GTP, 5 mg of bulk S.solfataricus tRNA, 2 ml of [ 35 S]methionine (1200 Ci/mmol at 10 mCi/ml) and $10 pmol of each mRNA. The mixtures were incubated at 70 C for 45 min. After this time, the synthesized proteins were resolved by electrophoresis 12.5% acrylamide-SDS gels and revealed by autoradiography of the dried gels on an Instant Imager apparatus.
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