EXPRESSION OF RECOMBINANT ACTIN 5C FROM DROSOPHILA IN THE METHYLOTROPHIC YEAST PICHIA PASTORIS
O. V. Nevzglyadova,1 A. V. Artemov,1 V. V. Zenin,1 V. V. Verkhusha,2
M. M. Shavlovsky,3 O. I. Povarova,1 Olesya V. Stepanenko,1 I. M. Kuznetsova,1
K. K. Turoverov 1
1 Institute of Cytology RAS, St. Petersburg, 2 Center for Molecular Medicine, Moscow State
University, and 3 Institute for Experimental Medicine RAMS, St. Petersburg;
e-mail: kkt@mail.cytspb.rssi.ru
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.
Key words: recombinant actin, green fluorescent protein, GFP-actin fusion, Pichia pastoris,
actin cytoskeleton
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