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      Overexpression, purification, and functional analysis of recombinant human tubulin dimer.

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          Abstract

          Microtubules consisting of tubulin dimers play essential roles in various cellular functions. Investigating the structure-function relationship of tubulin dimers requires a method to prepare sufficient quantities of recombinant tubulin. To this end, we simultaneously expressed human α1- and β3-tubulin using a baculovirus-insect cell expression system that enabled the purification of 5mg recombinant tubulin per litre of cell culture. The purified recombinant human tubulin could be polymerized into microtubules that glide on a kinesin-coated glass surface. The method provides a powerful tool for in vitro functional analyses of microtubules.

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          Author and article information

          Journal
          FEBS Lett.
          FEBS letters
          1873-3468
          0014-5793
          Nov 1 2013
          : 587
          : 21
          Affiliations
          [1 ] Laboratory for Molecular Biophysics, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
          Article
          S0014-5793(13)00661-3
          10.1016/j.febslet.2013.08.032
          24021646
          7af371ac-357d-4c5b-a2d7-4019fb52d624
          Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
          History

          Baculovirus-insect cell expression,Kinesin motility,MAP,PTM,Tubulin assembly,WT,microtubule-associated protein,posttranslational modification,wild-type

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