Overexpression, purification, and functional analysis of recombinant human tubulin dimer | Zendy

Minoura Itsushi | Zendy; Hachikubo You | Zendy; Yamakita Yoshihiko | Zendy; Takazaki Hiroko | Zendy; Ayukawa Rie | Zendy; Uchimura Seiichi | Zendy; Muto Etsuko | Zendy

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

Author(s) -

Minoura Itsushi,

Hachikubo You,

Yamakita Yoshihiko,

Takazaki Hiroko,

Ayukawa Rie,

Uchimura Seiichi,

Muto Etsuko

Publication year - 2013

Publication title -

febs letters

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.593

H-Index - 257

eISSN - 1873-3468

pISSN - 0014-5793

DOI - 10.1016/j.febslet.2013.08.032

Subject(s) - tubulin , recombinant dna , microtubule , dimer , microbiology and biotechnology , in vitro , function (biology) , cell culture , biology , microtubule associated protein , chemistry , biophysics , biochemistry , genetics , gene , organic chemistry

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 5 mg 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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