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Abstract
An in vitro DNA-directed protein-synthesizing system of Bacillus subtilis was developed using vegetative and sporulating cell extracts. Protease activity was inhibited by the addition of three kinds of protease inhibitors and removed from the extracts by hemoglobin-Sepharose treatment. Endogenous RNA synthesis was very low because of elimination of endogenous DNA by polyethylene glycol 6000 treatment of the supernatant. Protein synthesis was dependent on the DNA template, ribosomes and supernatant. When pUB110 DNA was used as a template, three proteins (Kl, K2 and K5) which have the same molecular weights as those synthesized in vivo were synthesized in vitro with vegetative, T2 (2 hr after the end of logarithmic growth) and T4 cell extracts.