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PMID: 2038331 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Characterization of recombination intermediates from DNA injected into Xenopus laevis oocytes: evidence for a nonconservative mechanism of homologous recombination.

Molecular and cellular biology ·Vol. 11 ·No. 6 ·1991-06-00 ·Pages 3278-87

Maryon E, Carroll D

Abstract

Homologous recombination between DNA molecules injected into Xenopus laevis oocyte nuclei is extremely efficient if injected molecules have overlapping homologous ends. Earlier work demonstrated that ends of linear molecules are degraded by a 5'----3' exonuclease activity, yielding 3' tails that participate in recombination. Here, we have characterized intermediates further advanced along the recombination pathway. The intermediates were identified by their unique electrophoretic and kinetic properties. Two-dimensional gel electrophoresis and hybridization with oligonucleotide probes showed that the intermediates had heteroduplex junctions within their homologous overlaps in which strands ending 3' were full length and those ending 5' were shortened. Additional characterization suggested that these intermediates had formed by the annealing of complementary 3' tails. Annealed junctions made in vitro were rapidly processed to products, indicating that they are on the normal recombination pathway. These results support a nonconservative, single-strand annealing mode of recombination. This recombination mechanism appears to be shared by many organisms, including bacteria, fungi, plants, and mammals.

MeSH Terms
Animals Base Composition DNA, Bacterial/administration & dosage,genetics DNA, Single-Stranded/genetics,isolation & purification Female Kinetics Microinjections Models, Genetic Molecular Weight Nucleic Acid Hybridization Oocytes/physiology Plasmids Recombination, Genetic Restriction Mapping Tetracycline Resistance/genetics Xenopus laevis
Chemicals
DNA, Bacterial DNA, Single-Stranded
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Maryon E
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City 84132.
Carroll D
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.11.6.3278-3287.1991
Published
1991-06-00
Pages
3278-87
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360180
Subset
IM
Grants
NIGMS NIH HHS · T32-GM07464 · United States
External Links
PubMed source DOI: 10.1128/mcb.11.6.3278-3287.1991 PMC Full Text View journal details
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