Nuclear Fraction of Bacillus subtilis as a Template for Ribonucleic Acid Synthesis

S. Mizuno, H. R. Whiteley
1968 Journal of Bacteriology  
A "nuclear fraction" prepared from Bacillus subtilis was a more efficient template than purified deoxyribonucleic acid for the synthesis of ribonucleic acid by exogenously added ribonucleic acid polymerase isolated from B. subtilis. The initial rate of synthesis with the nuclear fraction was higher and synthesis continued for several hours, yielding an amount of ribonucleic acid greater than the amount of deoxyribonucleic acid used as the template. The product was heterogenous in size, with a
more » ... s in size, with a large portion exceeding 23S. When purified deoxyribonucleic acid was the template, a more limited synthesis was observed with a predominantly 7S product. However, the ribonucleic acids produced in vitro from these templates were very similar to each other and to in vivo synthesized ribonucleic acid as determined by the competition of ribonucleic acid from whole cells in the annealing of in vitro synthesized ribonucleic acids to deoxyribonucleic acid. Treatment of the nuclear fraction with heat (60 C for 15 min) or trypsin reduced the capacity of the nuclear fraction to synthesize ribonucleic acid to the level observed with purified deoxyribonucleic acid. MATERIALS AND METHODS Cultivation of bacteria. B. subtilis strain 6, B. amyloliquefaciens ("strain K" obtained from B. Maruo, Institute of Applied Microbiology, University of Tokyo), and E. coli B were grown at 37 C with aeration in a medium containing: 0.1% Casamino Acids (Difco), 0.2% glucose (added as a sterile solution), 0.7% K2HP04, 0.3% KH2P04, 0.05% sodium citrate, 0.01% MgSO4-7H20, 0.1% (NH4)2S04, and 0.0002% L-tryptophan; the medium was adjusted to 1221 on May 8, 2020 by guest http://jb.asm.org/ Downloaded from 1222
doi:10.1128/jb.95.4.1221-1237.1968 fatcat:2ww27mq4sfctbnk2tpnlej6u54