Rapid purification of highly active ribosomes from Escherichia coli

Highly active salt-washed ribosomes from Escherichia coli are isolated by gel filtration on Sephacryl S-200 in a buffer containing 1 m ammonium chloride and putrescine, spermidine, calcium, and magnesium ions. Up to several hundred grams of cells can be processed in less than 24 h. The ribosome solu...

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Bibliographic Details
Published inAnalytical biochemistry Vol. 105; no. 1; pp. 369 - 374
Main Author Jelenc, Pierre C.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.01.1980
Subjects
Ammonium Chloride
Cell Fractionation - methods
Chromatography, Gel - methods
Escherichia coli - metabolism
Escherichia coli - ultrastructure
Ribosomes - metabolism
Ribosomes - ultrastructure
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Summary:Highly active salt-washed ribosomes from Escherichia coli are isolated by gel filtration on Sephacryl S-200 in a buffer containing 1 m ammonium chloride and putrescine, spermidine, calcium, and magnesium ions. Up to several hundred grams of cells can be processed in less than 24 h. The ribosome solution made 30% (v/v) in methanol is best stored as a liquid at −20°C. An improved poly(U)-dependent peptide synthesis assay is described in which phenylalanine polymerization is greatly enhanced while the misincorporation of leucine is kept at in vivo levels.
Bibliography:ObjectType-Article-1
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ISSN:0003-2697
1096-0309
DOI:10.1016/0003-2697(80)90472-8