Gamma tubulin and microtubule nucleation in mammalian cells

Abstract

Publisher Summary This chapter describes methods to study the function of γ-tubulin in mammalian cells. Antibody interference is a method often used to study the properties of proteins, using antibodies that react with the protein of interest and block its function. This approach can also be used to investigate the function of γ-tubulin in the nucleation of microtubule assembly in mammalian cells. The γ-tubulin antibody can be generated to conserved regions of this protein, and then the antibody can be incubated with permeabilized cells to study γ-tubulin function in vitro or injected into cultured mammalian cells to study γ-tubulin function in vivo. Expression of exogenous γ-tubulin is another important approach to study the function of γ-tubulin in microtubule nucleation in vivo in mammalian cells. To doso, a γ-tubulin expression vector with a strong promoter needs to be constructed and, on transfection, the promoter in the vector can efficiently drive the expression of γ-tubulin in mammalian cells. Consequently, γ-tubulin and its function in microtubule nucleation can be investigated.