Open AccessPreparation of a mRNA‐Dependent Cell‐Free Translation System from Whole Cells of Saccharomyces cerevisiae
Author(s) -
HOFBAUER Reinhold,
FESSL Friederike,
HAMILTON Barbara,
RUIS Helmut
Publication year - 1982
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1982.tb05867.x
Subject(s) - spheroplast , saccharomyces cerevisiae , yeast , micrococcal nuclease , cell free system , translation (biology) , messenger rna , biochemistry , protein biosynthesis , chemistry , biology , enzyme , microbiology and biotechnology , dna , gene , escherichia coli , chromatin , nucleosome
A cell‐free protein‐synthesizing system has been prepared from Saccharomyces cerevisiae by mechanical breakage of cells, isolation of a 30000xg supernatant fraction and removal of endogenous mRNA by treatment with micrococcal nuclease. The system thus isolated is dependent on added mRNA and translates yeast mRNA to discrete products, many of them identical with yeast proteins synthesized in vivo . Activity and properties of this system are comparable to those of other eukaryotic cell‐free translation systems. It offers the following advantages compared to yeast translation systems described previously. (a) Its isolation is simple and fast. (b) Since it is not isolated from spheroplasts there is no danger of its inactivation by contaminants in enzymes used for spheroplast preparation. (c) Isolation appears to be less strain‐dependent and can be carried out starting from cells in various physiological states.