Expression of recombinant actin 5C from Drosophila in the methylotrophyc yeast Pichia pastoris

O. V. Nevzglyadova, A. V. Artemov, V. V. Zenin, V. V. Verkhusha, M. M. Shavlovsky, O. I. Povarova, O. V. Stepanenko, I. M. Kuznetsova, K. K. Turoverov

Research output: Contribution to journalArticle

Abstract

A system of expression for the foreign actin gene in yeast cells Pichia pastoris has been developed. As a target protein, the Drosophila cytoplasmic actin 5C, which has 90% homology to the β-actin of higher eukaryotes, was used. In the present work, in order to develop conditions for biosynthesis of the target protein in yeast cells and a purification procedure for the recombinant protein, a GFP-actin fusion protein containing green fluorescent protein (GFP) as a fusion tag was expressed and purified. The size and survival of P. pastoris cells producing recombinant protein were characterized and shown to depend on the accumulation of recombinant actin. The purified fusion protein was used to obtain a polyclonal antibody necessary for testing for recombinant actin.

Original languageEnglish (US)
Pages (from-to)248-258
Number of pages11
JournalCell and Tissue Biology
Volume1
Issue number3
DOIs
StatePublished - Jun 1 2007

Keywords

  • GFP-actin fusion protein
  • Green fluorescent actin
  • Pichia pastoris actin cytoskeleton
  • Recombinant actin

ASJC Scopus subject areas

  • Cell Biology

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  • Cite this

    Nevzglyadova, O. V., Artemov, A. V., Zenin, V. V., Verkhusha, V. V., Shavlovsky, M. M., Povarova, O. I., Stepanenko, O. V., Kuznetsova, I. M., & Turoverov, K. K. (2007). Expression of recombinant actin 5C from Drosophila in the methylotrophyc yeast Pichia pastoris. Cell and Tissue Biology, 1(3), 248-258. https://doi.org/10.1134/S1990519X07030078