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Transgene elimination in genetically modified dry bean and soybean lines
Eduardo Romano1,2, Alexandre Soares1, Karina Proite1, Suzana Neiva1, Maíra Grossi1,
Josias C. Faria3, Elíbio L. Rech1 and Francisco J.L. Aragão1
1Embrapa Recursos Genéticos e Biotecnologia, Parque Estação Biológica, Final Av. W3 Norte,
70770-900 Brasília, DF, Brasil
2Departamento de Biologia Celular, Universidade de Brasília, Campus Universitário, Asa Norte,
70910-900 Brasília, DF, Brasil
3Embrapa Arroz e Feijão, Caixa Postal 179, 75375-000 Santo Antônio de Goiás, GO, Brasil
Corresponding author: E. Romano
E-mail: romano@cenargen.embrapa.br
Genet. Mol. Res. 4 (2): 177-184 (2005)
Received July 27, 2004
Accepted November 10, 2004
Published May 18, 2005

ABSTRACT. Transgene elimination is a poorly studied phenomenon in plants. We made genetic and molecular studies of a transgenic dry bean line immune to bean golden mosaic geminivirus and a soybean line. In both lines, the transgenes were stable during the vegetative phase but were eliminated during meiosis. Due to its potential biotechnological value, this transgenic line was micropropagated by grafting and the vegetative copies were studied for more than two years. More than 300 plants of progeny were obtained during this period, demonstrating that the phenomenon of elimination was consistently repeated and offering an opportunity for detailed study of transgene elimination, including the characterization of the integration sites. Cloning and sequencing of the transgenic loci, reciprocal crosses to untransformed plants, genomic DNA blots, and GUS assays were performed in the transgenic lines. Based on the molecular and genetic characterization, possible mechanisms involved in transgene elimination include intrachromosomal recombination, genetic instability resulting from the tissue culture manipulations, and co-elimination of transgenes, triggered by a process of genome defense.

Key words: Transgene locus structure, Gene integration, Biolistics, Transgene elimination

 

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