A REQUIREMENT FOR GENE-SPECIFIC DEOXYRIBONUCLEIC ACID FOR THE CELL-FREE SYNTHESIS OF β-GALACTOSIDASE

Abstract

By disruption of whole cells of Escherichia coli in the French pressure cell, a system can be prepared that carries out the cell-free synthesis of [beta]-galactosidase. The system must be prepared from pre-induced cells and requires a particle fraction sedimenting at 105,000 X g, a supernatant fraction, a source of energy, the nucleoside di- and triphosphates, and the inducer. The system is also capable of a vigorous incorporation of C14 leucine into protein and much of the radioactive protein can be precipitated with anti-[beta]-galactosidase serium. Both enzyme synthesis and incorporation of labeled amino acid are sensitive to ribonuclease, chloramphenicol, and deoxyribo-nuclease.