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A cell-free protein-synthesizing system prepared from mouse sarcoma 180 was characterized by use of RNA from mengo virus and sea urchin egg. In the presence of exogenous mammalian transfer RNA, total sea urchin egg RNA and mengo RNA direct incorporation of [(3)H]leucine into acid-insoluble material. The system is extremely efficient in that a stimulation of 100-times over background can be obtained. Studies with formylmethionyl-transfer RNA, as well as with inhibitors of initiation, indicate that multiple initiation occurs; further, 85-90% of all chains made in vitro are subsequently released from ribosomes. An average translation time of 3.5 min was determined with messenger RNA of sea urchin egg, and product analysis indicates that high-molecular-weight products (greater than 50,000 molecular weight) are being made in vitro. Sequences of sea urchin egg RNA containing poly(A) act as messenger RNA.
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