In vitro system from Escherichia coli that catalyzes generalized genetic recombination

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This paper reports an in vitro system for studying generalized genetic recombination. The system uses extracts from Escherichia coli as a source of enzymes and plasmid DNA molecules as substrates. Unit-size plasmid DNA rings are converted into genomes fused at a region of DNA homology at a frequency of about 5-10% over a period of hours. That the fused structures are the result of recombination is supported by two lines of evidence. When two partially homologous plasmids of different sizes are used as substrates for the in vitro system, intermediates containing one plasmid of each size are obtained. Furthermore, fused structures are not formed with high efficiency in extracts from recombination-deficient (Rec A-) cells.

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