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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Dujon, B. Puchta, H. Hohn, B. |
| Description | Author Affiliation: Puchta H ( Friedrich Miescher-Institut, Basel, Switzerland.); |
| Abstract | Genomic double-strand breaks (DSBs) are key intermediates in recombination reactions of living organisms. We studied the repair of genomic DSBs by homologous sequences in plants. Tobacco plants containing a site for the highly specific restriction enzyme I-Sce I were cotransformed with Agrobacterium strains carrying sequences homologous to the transgene locus and, separately, containing the gene coding for the enzyme. We show that the induction of a DSB can increase the frequency of homologous recombination at a specific locus by up to two orders of magnitude. Analysis of the recombination products demonstrates that a DSB can be repaired via homologous recombination by at least two different but related pathways. In the major pathway, homologies on both sides of the DSB are used, analogous to the conservative DSB repair model originally proposed for meiotic recombination in yeast. Homologous recombination of the minor pathway is restricted to one side of the DSB as described by the nonconservative one-sided invasion model. The sequence of the recombination partners was absolutely conserved in two cases, whereas in a third case, a deletion of 14 bp had occurred, probably due to DNA polymerase slippage during the copy process. The induction of DSB breaks to enhance homologous recombination can be applied for a variety of approaches of plant genome manipulation. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 10 |
| Volume Number | 93 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1996-07-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | DNA Repair Genetics Plants Recombination, Genetic Cloning, Molecular DNA Damage DNA, Plant Metabolism Deoxyribonucleases, Type II Site-Specific Gene Targeting Genome, Plant Models, Genetic Molecular Sequence Data Microbiology Plants, Genetically Modified Plants, Toxic Polymerase Chain Reaction Rhizobium Saccharomyces Cerevisiae Proteins Sequence Deletion Tobacco Transformation, Genetic Comparative Study Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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