A COMPARATIVE LEVEL OF EXPRESSION OF SOME PROTEINS IN XL2 CELL SYNCHRONIZED ON DIFFERENT
PHASES OF CELL CYCLE
R. E. Uzbekov
Divisioin of Electron Microscopy, A. N. Bclozcrsky Institute of Physico-Chemical Biology,
Moscow State University, Russia;
e-mail: rustuzbekov@aol.com
Cells of cultured line XL2 (Xenopus laevis) were synchronized by a combine effect of serum deprivation,
aphidicolin, nocodazole and ALLN treatments. Four fractions were prepared, with maximum percentage of cells being in G1,
S and G2 phases of cell cycle, and in mitosis, respectively. Comparative levels of six different proteins (beta-tubulin,
DNA topoisomerase IIα, Xenopus Aurora A kinase pEg2, kinesin-like motor protein XlEgS, and two members of condensis
family proteins pEg7 (XCAP D2) and XCAP E were detected by quantitative Western blot analysis of these fractions. We used a new
method of mathematic processing of data that commonly provides a possibility to calculate a comparative quantity of proteins in
hypothetically "clean" fraction composed of cells being in the same phase of the cell cycle. This method makes it possible to use even
partly synchronized cell cultures for analysis of changes in protein quantity, provided a precide determination of cell population
composition is made.
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