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Chapter 7 Isolation of Vacuoles from Yeasts

1975; Elsevier BV; Linguagem: Inglês

10.1016/s0091-679x(08)60954-1

0091-679X

Andres Wiemken,

Photosynthetic Processes and Mechanisms

This chapter discusses the isolation of vacuoles from yeasts. Vacuoles are components of an elaborate membrane system that also includes the endoplasmic reticulum and the Golgi complex. Because the components of this system have manifold interconnections and are involved in a continuous process of interconversion, it is not always possible to distinguish clearly among them. In the chapter, methods are described by which vacuoles can be isolated from yeast cells. All these methods involve an initial transformation of the yeast cells into spheroplasts. The spheroplasts contain only one or two large vacuoles, whereas a multitude of small vacuoles is often present in growing yeast cells. This facilitates isolation, because the large vacuoles can easily be identified throughout the entire isolation procedure using a light microscope. It, however, limits the validity of the results obtained from studies on these vacuoles. In the chapter, extraction of intact vacuoles from spheroplasts and characterization of isolated vacuoles are describes.