Expression of recombinant actin 5C from drosophila in the methylotrophic yeast pichia pastoris

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

Research output: Contribution to journalArticle

Abstract

A system for actin expression in cells of yeast Pichia pastoris was constructed. Drosophila actin 5C, by 90% homologous to β-actin of higher eukaryotes, was used as a target protein. To improve the procedures of target protein biosynthesis in yeast cells and of extraction and purification of recombinant actin the fusion protein GFP-actin 5C, having fluorescence protein GFP as a reporter part, was expressed and purified. The dimensions and resistance of yeast cells producing recombinant actin were characterized. It was shown that the size and form of cells depended on the accumulation of recombinant protein. The purified fusion protein was used for obtaining polyclonal antibody for testing recombinant actin.

Original languageEnglish (US)
Pages (from-to)300-310
Number of pages11
JournalTsitologiya
Volume49
Issue number4
Publication statusPublished - Jul 6 2007
Externally publishedYes

    Fingerprint

Keywords

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

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology

Cite this

Nevzglyadova, O. V., Artemov, A. V., Zenin, V. V., Verkhusha, V. V., Shavlovsky, M. M., Povarova, O. I., ... Turoverov, K. K. (2007). Expression of recombinant actin 5C from drosophila in the methylotrophic yeast pichia pastoris. Tsitologiya, 49(4), 300-310.