Mutations produced by DNA polymerase III holoenzyme of Escherichia coli after in vitro synthesis in the absence of single-strand binding protein
Department of Biochemistry and Molecular Pharmacology
Base Sequence; Chromosome Deletion; DNA Polymerase III; *DNA Replication; DNA Transposable Elements; DNA, Bacterial; DNA, Single-Stranded; Escherichia coli; Genes, Bacterial; Molecular Sequence Data; Mutation; Plasmids; Templates, Genetic; Tetracycline Resistance
Biochemistry, Biophysics, and Structural Biology | Pharmacology, Toxicology and Environmental Health
Single-stranded plasmid DNA, containing the mnt gene, was replicated in vitro with DNA polymerase III holoenzyme. Escherichia coli mutH bacteria, defective in mismatch repair, were transformed with the products of in vitro synthesis. Mutations in mnt were readily identified and 33 out of 65 isolates were single base changes including transition, transversion and frameshift mutations. The remaining 32 isolates were deletions of apparently random length and substitutions (deletion/insertions). The intergenic deletions as well as the transition and frameshift mutations were identical to those previously isolated from mismatch repair-defective cells in vivo.
Mol Microbiol. 1990 Oct;4(10):1645-52.
Carraway M, Rewinski C, Marinus MG. (1990). Mutations produced by DNA polymerase III holoenzyme of Escherichia coli after in vitro synthesis in the absence of single-strand binding protein. Biochemistry and Molecular Biotechnology Publications. Retrieved from https://escholarship.umassmed.edu/bmp_pp/35