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J Mol Biol ; 291(1): 29-46, 1999 Aug 06.
Article in English | MEDLINE | ID: mdl-10438604

ABSTRACT

Illegitimate recombination is the prevailing molecular mechanism for the integration of recombinant DNA into the genome of most eukaryotic systems and the generation of deletions by intrachromosomal recombination. We developed a ?selectable marker system to screen for intrachromosomal illegitimate recombination events in order to assess the sequence and structure-specific requirements for illegitimate recombination in tobacco. In 12 illegitimate recombination products analysed, we found that all deletion termini localise to sites of palindromic structures or to A+T-rich DNA elements. All deletion termini showed microhomologies of two to six nucleotides. In three plants, the recombination products contained filler-DNA or an inversion of an endogenous segment. Our data strongly suggest that illegitimate recombination in plants is mediated by a DNA synthesis-dependent process, and that this mechanism is promoted by DNA regions that can form palindromic structures or facilitate DNA unwinding.


Subject(s)
DNA, Plant/genetics , Nicotiana/genetics , Plants, Toxic , Recombination, Genetic , Adenosine/chemistry , Base Pairing , Base Sequence , DNA, Plant/analysis , DNA, Plant/chemistry , Molecular Sequence Data , Nucleic Acid Conformation , Polymerase Chain Reaction , Protoplasts/physiology , Sequence Homology, Nucleic Acid , Thymidine/chemistry
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