The Isolation of Tubulin and Actin from Mouse 3T3 Cells Transformed by Simian Virus 40 (SV3T3 Cells), an Established Cell Line Growing in Culture

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Abstract
Tubulin can be purified from mouse SV3T3 cells (3T3 cells transformed by SV40 virus) by several cycles of temperature-dependent polymerization and depolymerization. EM analysis of the final product reveals morphologically normal microtubules. Homogeneous actin can be isolated as a byproduct of the purification procedure. Mouse SV3T3 actin and skeletal muscle actin were compared by fingerprint analysis of the tryptic peptides obtained from performic-acid-oxidized protein. The 2 actins show a high degree of homology although apparently 5 of the 25 spots visualized by fluorescamine show a difference in chromatographic mobility. The purification procedure described allows the rapid isolation of actin and tubulin from tissue culture cells in sufficient amounts for comparative biochemical studies.