Abstract
Summary
A cell-free system which was active with respect to amino acid-14C incorporation into hot TCA insoluble material was derived from Cellvibrio gilvus and characterized. This amino-acid incorporation was typical in its requirement for ribosomes, pH 5 fraction, ATP, and magnesium ions, and in its inhibition by ribonuclease and puromycin. Deoxyribonuclease and actinomycin D had little effect, whereas chloramphenicol caused 65% inhibition. The optimal condition for incorporation was 30 min incubation at 30° and at pH 8.0. The addition of an exogenous source of a polyamine (spermidine, spermine, or putrescine) had little effect on this system. Poly-U dependent phenylalanine incorporation was observed with this system and found to be stimulated by an exogenous source of tRNA, although tRNA had no stimulatory effect on leucine incorporation.
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