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      Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase

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      Gene
      Elsevier BV

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          Abstract

          <p class="first" id="d9120341e61">Plasmid expression vectors have been constructed that direct the synthesis of foreign polypeptides in Escherichia coli as fusions with the C terminus of Sj26, a 26-kDa glutathione S-transferase (GST; EC 2.5.1.18) encoded by the parasitic helminth Schistosoma japonicum. In the majority of cases, fusion proteins are soluble in aqueous solutions and can be purified from crude bacterial lysates under non-denaturing conditions by affinity chromatography on immobilised glutathione. Using batch wash procedures several fusion proteins can be purified in parallel in under 2 h with yields of up to 15 micrograms protein/ml of culture. The vectors have been engineered so that the GST carrier can be cleaved from fusion proteins by digestion with site-specific proteases such as thrombin or blood coagulation factor Xa, following which, the carrier and any uncleaved fusion protein can be removed by absorption on glutathione-agarose. This system has been used successfully for the expression and purification of more than 30 different eukaryotic polypeptides. </p>

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          Author and article information

          Journal
          Gene
          Gene
          Elsevier BV
          03781119
          July 1988
          July 1988
          : 67
          : 1
          : 31-40
          Article
          10.1016/0378-1119(88)90005-4
          3047011
          b118f455-1911-4a5b-8431-c0e5801adced
          © 1988

          http://www.elsevier.com/tdm/userlicense/1.0/

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