To proceed the works on induced mutagenesis in plasmids, mutagenic effects of chemicals on the DNA of RSF2124 plasmid mediating colicine E1 biosynthesis and resistance to ampicillin, were studied. After exposure to mutagens, plasmid DNA was used to transform Escherichia coli C600 rk-mk-cells. The lethal effect was estimated from inactivation of the ampicillin marker, the mutagenic effect being measured by the appearance of mutants unable to synthesize colicine (Col-).
View Article and Find Full Text PDFMol Gen Mikrobiol Virusol
August 1985
The technique of localized in vitro mutagenesis in the cohesive ends of plasmid pBR322 DNA has been elaborated (separately for BamHI and HindIII sites). Plasmid DNA digested by restriction endonucleases has been treated with sodium bisulphite deaminating cytosine to form uracil in single stranded DNA (cohesive ends of the plasmid). The mutagenized plasmid DNA, free of mutagen, has been treated with bacteriophage T4 ligase.
View Article and Find Full Text PDFThis paper describes the results of treating plasmid DNA in vitro with mutagens, to obtain mutations both in plasmid genes and chromosomal genes comprised within the plasmid, thus avoiding disorganization characteristic of in vivo mutagenesis. The model system is represented by DNA of RSF2124 responsible for colicine E1 synthesis and resistance to ampicillin. Col- mutants were looked for after exposure to UV- and gamma-irradiation.
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