In vitro Protein Synthesis and Measurement of the Stability of Messenger RNA in the Blue-green Alga, Anabaena variabilis

Abstract

A cell-free preparation of the blue-green alga, Anabaena variabilis, that incorporates 14C-labelled amino acids into protein has been prepared and characterized. The activation of amino acids to amino acid transfer RNAs was characterized, and assembly of these into peptides has been described with respect to cofactor requirements and antibiotic sensitivity. The properties of these systems and the effect of antibiotics on them are similar to those of bacteria. Both natural and synthetic messenger RNA were effective in peptide formation. The kinetics of incorporation of [14C]uracil into RNA has been examined and the stability of labelled RNA from A. variabilis measured by radioactivity loss and by its role in directing peptide synthesis. The half-life of messenger RNA from this organism is approximately 12 min, and as such is comparable to that of bacteria when based upon the mean generation time of the respective organisms.