Poster: Emerging technologies

Abs # 909: Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants

Presenter:	Park, Jeongmoo , jmpark@kaist.ac.kr
Authors	Park, Jeongmoo  (A)   Lee, YoungKoung  (A)   Hong, Seho  (A)   Chung, Wonil  (A)
Affiliations:	(A): Dept. of Biol. Sci., KAIST

A negative selectable marker gene, codA, was successfully co-transformed with 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 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 R1 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.