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J Bacteriol. 1974 June; 118(3): 974-979
Copyright © 1974 American Society for Microbiology. All Rights Reserved.

Tests for Deoxyribonucleic Acid Synthesis in Toluenized Bacillus subtilis Cells

^a Department of Biochemical Sciences, Princeton University, Princeton, New Jersey 08540

ABSTRACT

Several tests were devised to further characterize deoxyribonucleic acid (DNA) synthesis in toluenized Bacillus subtilis cells. Vigorous agitation of toluenized cells (localization test) demonstrated that the DNA replication is exclusively a cell-associated process. A DNA "repair" condition was also applied to toluenized cells and shown to be distinct from DNA replication in its DNA polymerase I dependency and its ability to synthesize DNA on template which is either cell associated or free, outside the cell. This repair condition was used in conjunction with the localization test to demonstrate the penetration of deoxyribonuclease I and possibly DNA polymerase I into toluenized cells. Therefore, we suggest that the localization test can be used to test the penetration of proteins into toluenized cells for both the DNA repair and replication processes.

¹ Present address: Division of Biological and Medical Research, Argonne National Laboratory, Argonne, Ill. 60439.

² Present address: Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, Colo. 80302.