Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants

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A negative selectable marker gene, codA, was successfully co-transformed with a GUS reporter gene to develop selectable marker gene-free transgenic plants. The pNC binary vector contained a T-DNA harboring the codA gene next to the nptII gene, while a second binary vector, pHG, contained a GUS reporter gene. Tobacco plants (Nicotiana tabacum cv. Samsun NN) were co-transformed via the mixture method with Agrobacterium tumefaciens LBA4404 strains harboring pNC and pHG, respectively. Seeds harvested from the co-transformants were sown on germination media containing 5-fluorocytosine (5-FC). Analysis of the progeny by GUS staining and PCR amplification revealed that all of the 5-FC-resistant R-1 plants were codA free, and that the codA gene segregated independently of the GUS gene. Because codA-free seedlings developed normally on 5-FC-containing medium, we suggest that co-transformation with negatively selectable markers is a viable method for the production of easily distinguished, selectable marker gene-free transgenic plants.
Publisher
Springer
Issue Date
2004-09
Language
English
Article Type
Article
Keywords

T-DNAS; AGROBACTERIUM-TUMEFACIENS; VECTOR SYSTEM; TRANSFORMATION; SELECTION; CYTOCHROME-P450; SEGREGATION; EXPRESSION; HERBICIDE; CELL

Citation

THEORETICAL AND APPLIED GENETICS, v.109, no.8, pp.1562 - 1567

ISSN
0040-5752
DOI
10.1007/s00122-004-1790-x
URI
http://hdl.handle.net/10203/82784
Appears in Collection
BS-Journal Papers(저널논문)
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