Gene targeting in plants
Paszkowski, J.; Baur, M.; Bogucki, A.; Potrykus, I.
EMBO Journal 7(13): 4021-4026
1988
ISSN/ISBN: 0261-4189 PMID: 16453864 Document Number: 324913
Although the generation of transgenic plants is now routine, the integration of foreign genetic information has so far been at random sites in the genome. We now present evidence for directed integration into a predicted location in the host plant genome. Protoplasts of transgenic tobacco (Nicotiana tabacum) plants carrying copies of a partial, non-functional drug-resistance gene in the nuclear DNA were used as recipients for DNA molecules containing the missing part of the gene. Molecular and genetic data confirm the integration of the foreign DNA through homologous recombination within overlapping parts of the protein coding region, resulting in the formation of an active gene in the host chromosome. This approach is referred to as gene targeting. The gene targeting frequency (the number of drug-resistant clones resulting from gene correction compared to the number of resistant clones from parallel experiments with a similar non-interrupted hybrid gene) was 0.5-4.2 .times. 10-4. These experiments demonstrate the possibility of producing transgenic plants with desired modifications to a specific nuclear gene.