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http://dx.doi.org/10.5010/JPB.2013.40.4.198

Development of transgenic cucumbers expressing Arabidopsis Nit gene  

Jang, Hyun A (Plant Systems Engineering Research Center, KRIBB)
Lim, Ka Min (Department of Oriental Pharmaceutical Development, Nambu University)
Kim, Hyun A (Plant Systems Engineering Research Center, KRIBB)
Park, Yeon-Il (Department of Biology, Chungnam National Univ.)
Kwon, Suk Yoon (Plant Systems Engineering Research Center, KRIBB)
Choi, Pil Son (Department of Oriental Pharmaceutical Development, Nambu University)
Publication Information
Journal of Plant Biotechnology / v.40, no.4, 2013 , pp. 198-202 More about this Journal
Abstract
To produce transgenic cucumber expressing Nit gene coffering abiotic resistance, the cotyledonary-node explants of cucumber (cv. Eunsung) were inoculated with A. tumefaciens transformed with pPZP211 or pCAMBIA2300 carrying Nit gene, that has cis-acting element involved in resistance to various abiotic environmental stresses. After co-cultivation, the procedures of selection, shoot initiation, shoot elongation, and plant regeneration were followed by cotyledonary-node transformation method (CTM, Jang et al. 2011). The putative transgenic plants were selected when shoots were grown to a length greater than 3 cm from the cotyledonary-node explants on selection medium supplemented with 100 mg/L paromomycin as a selectable agent. The confirmation of transgenic cucumber was based on the genomic PCR, Southern blot analysis, RT-PCR, and Northern blot analysis. A 105 shoots (4.12%) selected from the selection mediums were obtained from 2,547 explants inoculated. Of them, putative transgenic plants were only confirmed with 45 plants (1.77%) by genomic PCR analysis. Transgenic plants showed that the Nit genes integrated into each genome of 39 plants (1.53%) by Southern blot analysis, and the expression of gene integrated into cucumber genome was only confirmed at 6 plants (0.24%) by RT-PCR and Northern blot analysis. These results lead us to speculate that the genes were successfully integrated and expressed in each genome of transgenic cucumber.
Keywords
Abiotic gene; Agrobacterium; Nit gene; Transgenic cucumber;
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