Recombination of DNAs in Xenopus oocytes based on short homologous overlaps

Abstract

Linear molecules of pBR322 and closely related plasmid DNAs were injected into Xenopus oocyte nuclei. Such molecules were degraded unless their ends were recombined. Non-homologous ends were joined rarely, if at all, but measurable recombination was supported by homologous sequences of less than 10 base pairs (bp). The efficiency of recombination increased as the length and degree of homology improved, in the range of about 8-20 bp. The homologous sequences had to be very close to the original molecular ends (within about 20 bp); internal homologies, even when they included better matches, were never used. These observations are best accommodated by a model of recombination which envisions exonucleolytic resection to expose homologous sequences, followed by annealing of single-stranded tails, tidying up and sealing of the new joint. Some of the recombined plasmids had novel tetracycline resistance genes; their properties give some insight into the function of the tet gene product.