Efficient homologous recombination of linear DNA substrates after injection into Xenopus laevis oocytes.

AUTOR(ES)

Carroll, D

RESUMO

When DNA molecules are injected into Xenopus oocyte nuclei, they can recombine with each other. With bacteriophage lambda DNAs, it was shown that this recombination is stimulated greatly by introduction of double-strand breaks into the substrates and is dependent on homologous overlaps in the recombination interval. With plasmid DNAs it was shown that little or no recombination occurs between circular molecules but both intra- and intermolecular events take place very efficiently with linear molecules. As with the lambda substrates, homology was required to support recombination; no simple joining of ends was observed. Blockage of DNA ends with nonhomologous sequences interfered with recombination, indicating that ends are used directly to initiate homologous interactions. These observations are combined to evaluate possible models of recombination in the oocytes. Because each oocyte is capable of recombining nanogram quantities of linear DNA, this system offers exceptional opportunities for detailed molecular analysis of the recombination process in a higher organism.

Documentos Relacionados

• Effect of terminal nonhomologies on homologous recombination in Xenopus laevis oocytes.
• Intramolecular recombination between partially homologous sequences in Escherichia coli and Xenopus laevis oocytes.
• Accurate and efficient transcription of human c-myc genes injected into Xenopus laevis oocytes.
• Characterization of recombination intermediates from DNA injected into Xenopus laevis oocytes: evidence for a nonconservative mechanism of homologous recombination.
• Transcription of cloned Moloney murine leukemia proviral DNA injected into Xenopus laevis oocytes.