[果蝇重组肌动蛋白5C在甲基营养型酵母毕赤酵母中的表达]

[Expression of recombinant actin 5C from Drosophila in the methylotrophic yeast Pichia pastoris].

作者信息

Nevzgliadova O V, Artemov A V, Zenin V V, Verkhusha V V, Shavlovskiĭ M M, Povarova O I, Stepanenko O V, Kuznetsova I M, Turoverov K K

出版信息

Tsitologiia. 2007;49(4):300-10.

PMID:17657944

Abstract

A system for actin expression in cells of yeast Pichia pastoris was constructed. Drosophila actin 5C, by 90% homologous to beta-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.

摘要

构建了一种用于在巴斯德毕赤酵母细胞中表达肌动蛋白的系统。将与高等真核生物的β-肌动蛋白具有90%同源性的果蝇肌动蛋白5C用作靶蛋白。为了改进酵母细胞中靶蛋白生物合成以及重组肌动蛋白提取和纯化的程序，表达并纯化了具有荧光蛋白GFP作为报告部分的融合蛋白GFP-肌动蛋白5C。对产生重组肌动蛋白的酵母细胞的尺寸和抗性进行了表征。结果表明，细胞的大小和形态取决于重组蛋白的积累。纯化的融合蛋白用于制备多克隆抗体以检测重组肌动蛋白。