Illegitimate recombination in Xenopus: characterization of end-joined junctions

When linear DNAs are injected into Xenopus laevis eggs, they are converted Into several different kinds of recombination products. Some molecules undergo homologous recombination by a resection-annealing mechanism; some ends are precisely llgated; and some ends are Joined by illegitimate means. The...

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Bibliographic Details
Published in:Nucleic acids research 1994-02, Vol.22 (3), p.434-442
Main Authors: Lehman, Chris W., Trautman, Jonathan K., Carroll, Dana
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biological and medical sciences
Cell Nucleus - metabolism
Cell-Free System
DNA Primers - chemistry
Fundamental and applied biological sciences. Psychology
Genic rearrangement. Recombination. Transposable element
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Oocytes
Oogenesis
Ovum
Recombination, Genetic
Sequence Homology, Nucleic Acid
Xenopus
Xenopus laevis
Xenopus laevis - genetics
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Summary:When linear DNAs are injected into Xenopus laevis eggs, they are converted Into several different kinds of recombination products. Some molecules undergo homologous recombination by a resection-annealing mechanism; some ends are precisely llgated; and some ends are Joined by illegitimate means. The homologous and illegitimate products are also generated in nuclear extracts from stage VI Xenopus oocytes. In order to gain insight Into the mechanism(s) of illegitimate end joining, we amplified, cloned and sequenced a number of junctions from eggs and from oocyte extracts. The egg junctions fell into three categories: some with no homology at the join point that may have been produced by blunt-end ligation; some based on small, but significant homologies (5–10 bp); and some with matches of only 1 or 2 nucleotides at the joint. Junctions made in oocyte extracts were largely of the latter type. In the extracts, formation of illegitimate joints required the addition of all four deoxyribo-nucleoside triphosphates and was inhibited by aphidl-colin. This indicates that this process involves DNA synthesis, and mechanisms incorporating this feature are considered. The spectrum of recombination products formed in Xenopus eggs is very reminiscent of those produced from DNA introduced into mammalian cells.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/22.3.434