Total reconstitution of active small ribosomal subunits of the extreme halophilic archaeon Haloferax mediterranei |
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| Institution: | 1. Specialist Neonatal and Paediatric Surgery Department, Great Ormond Street Hospital, London, U.K.;2. Department of Radiology, Great Ormond Street Hospital, London, U.K.;1. Department of Earth, Environmental and Planetary Sciences, Brown University, Providence, RI 02912, USA;2. British Geological Survey, Nicker Hill, Keyworth NG12 5GG, UK;3. Centro de Astrobiologia (INTA-CSIC), Ctra Ajalvir km 4, Torrejon de Ardoz 28850, Spain;4. Department of Organismic and Evolutionary Biology, Harvard University, Çambridge, MA 02138, USA |
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| Abstract: | The small ribosomal subunit of the halophilic archaeon Haloferax mediterranei has been reconstituted from its dissociated rRNA and protein components. Efficient reconstitution of particles, fully active in poly(U)-dependent polyphenylalanine synthesis, occurs after 2 h of incubation at 36°C in the presence of l.5 M of (NH4)2SO4 100 mM of MgAc2, 20 mM Tris-HCI (pH 8.2) and 6 mM 2-mercaptoethanol. Important differences in the optimal ionic conditions for the reconstitution of the 30S and the 50S ribosomal subunits from Haloferax mediterranei have been found. K+ and NH4+ ions have differing abilities to promote the reconstitution of the particles. The assembly of 30S ribosomal subunits of H. mediterranei has a higher tolerance to ionic strength than the assembly of the 50S subunits and it is independent of the Mg2+concentration present in the system. |
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