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GrainGenes Reference Report: TPJ-32-433

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Reference	TPJ-32-433
Title	Complex transgene locus structures implicate multiple mechanisms for plant transgene rearrangement
Journal	Plant Journal
Year	2002
Volume	32
Pages	433-445
Author	Svitashev SK Pawlowski WP Makarevitch I Plank DW Somers DA
Abstract	Summary: To more fully characterize the internal structure of transgene loci and to gain further understanding of mechanisms of transgene locus formation, we sequenced more than 160 kb of complex transgene loci in two unrelated transgenic oat (Avena sativa L.) lines transformed using microprojectile bombardment. The transgene locus sequences from both lines exhibited extreme scrambling of non-contiguous transgene and genomic fragments recombined via illegitimate recombination. A perfect direct repeat of the delivered DNA, and inverted and imperfect direct repeats were detected in the same transgene locus indicating that homologous recombination and synthesis-dependent mechanism(s), respectively, were also involved in transgene locus rearrangement. The most unexpected result was the small size of the fragments of delivered and genomic DNA incorporated into the transgene loci via illegitimate recombination; 50 of the 82 delivered DNA fragments were shorter than 200 bp. Eleven transgene and genomic fragments were shorter than the DNA lengths required for Ku-mediated non-homologous end joining. Detection of these small fragments provided evidence that illegitimate recombination was most likely mediated by a synthesis-dependent strand-annealing mechanism that resulted in transgene scrambling. Taken together, these results indicate that transgene locus formation involves the concerted action of several DNA break-repair mechanisms
External Databases	Pubmed: 12445116
Keyword	[ Hide all but 1 of 25 ] avena avena sativa chromosome rearrangements detection dna double-strand break double-strand break (dsb) repair fragments genetic engineering homologous recombination illegitimate recombination junction regions mammalian cells microprojectile bombardment multiple non-homologous end joining (nhej) oat recombination repair repeat sequence t-dna integration transformation frequency transgene locus structure transgenic

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