Development and optimization of a coupled cell-free system for the synthesis of the transmembrane domain of the receptor tyrosine kinase ErbB3 |
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Authors: | N F Khabibullina E N Lyukmanova G S Kopeina Z O Shenkarev A S Arsen’ev D A Dolgikh M P Kirpichnikov |
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Institution: | 1.Moscow State University,Moscow,Russia;2.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry,Russian Academy of Sciences,Moscow,Russia |
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Abstract: | A coupled cell-free expression system (CECF) for the production of the transmembrane domain of the human receptor tyrosine
kinase ErbB3 (residues from 632 to 675) has been developed based on the Escherichia coli S30 extract. The synthesis of the domain in the soluble form in the presence of various detergents and in the form of an
insoluble precipitate of the reaction mixture has been examined. The conditions for the purification of the recombinant domain
obtained using the two approaches have been determined. The final yield of the target protein under optimal conditions was
1.8–2.0 mg per 1 ml of the reaction mixture. |
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