Reference: Warner JR and Gorenstein C (1978) The ribosomal proteins of Saccharomyces cerevisiae. Methods Cell Biol 20:45-60

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Abstract


The chapter studies the ribosomal proteins of Succharomyces cerevisiae because it is possible to obtain mutants which are likely to involve ribosomal proteins and focuses on the coordinated regulation of their synthesis. This chapter describes methods of preparing and analyzing the ribosomal proteins of S. cerevisiae and points out certain properties that are shared by the ribosomal proteins of all eukaryotes. Ribosomal proteins are analyzed directly on one-dimensional sodium dodecyl sulfate (SDS) gels without removing RNA or on two systems of two-dimensional polyacrylamide gel. The first is based on the Kaltschmidt???Wittman system and employs 6 M urea at pH 8.6 in the first dimension and at pH 4.5 in the second dimension. The second is a modified version of the Mets and Bogorad system and employs 8 M urea at pH 5.0 in the first dimension and sodium SDS in the second dimension. The two systems are described separately, and the results then compared. For both gel systems the importance of using proteins free of RNA cannot be overemphasized. RNA binds to ribosomal proteins even in urea solutions and causes poor resolution and decreased yields.

Reference Type
Journal Article | Research Support, U.S. Gov't, P.H.S.
Authors
Warner JR, Gorenstein C
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