Polynucleotide-dependent Incorporation of Amino Acids in a Cell-free System from Thermophilic Bacteria

From the ¹ From the Department of Biochemistry, New York University School of Medicine, New York, New York 10016

The preparation of a cell-free amino acid incorporation system from Bacillus stearothermophilus active at 65° and the conditions for optimal activity with synthetic polynucleotide messengers are described. The activity of this system is comparable to that of the conventional Escherichia coli system. Its thermal stability, high activity, and low nuclease content are convenient features for studies of protein synthesis and coding. Thus, a 1.4:1 copolymer of adenylic and uridylic acids, which, because of a high degree of ordered structure, is almost inactive in the E. coli system at 37°, promoted significant incorporation of isoleucine, leucine, and lysine in the B. stearothermophilus system at 65°.

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