Toluene treatment kills ceils of Escherichia coli and makes them permeable to molecules of low molecular weight. Despite the fact that these cells are no longer viable, they retain a number of their physiological functions and the ability to synthesize DNA, if supplied with the necessary substrates. This chapter describes the assay of DNA synthesis in toluene-treated cells based upon the incorporation of radioactively labeled deoxynucleotide residues into acid-insoluble polydeoxynucleotides. DNA synthesis in toluene-treated cells occurs in a normal manner if the reaction mixture contains Triton X-100, a nonionic detergent. One of the convincing arguments that DNA synthesis in the toluene-treated cell system duplicates active replication in vivo has been the temperature effect on temperature-sensitive replication mutants in vitro.
ASJC Scopus subject areas
- Molecular Biology