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http://dx.doi.org/10.5010/JPB.2007.34.4.375

Expression of CP4 5-Enol-Pyruvylshikimate-3- Phosphate Synthase Transgene in Inbred Line of Korean Domestic Maize (Zea may L.)  

Cho, Mi-Ae (Department of Oriental Pharmaceutical Development, Nambu University)
Kwon, Suk-Yoon (Korea Research Institute of Bioscience and Biotechnology (KRIBB))
Kim, Jin-Seog (Korea Research Institute of Chemical Technology (KRICT))
Lee, Byoung-Kyu (National Institute Crop Science)
Moon, Choo-Yeun (Kyung Woon University)
Choi, Pil-Son (Department of Oriental Pharmaceutical Development, Nambu University)
Publication Information
Journal of Plant Biotechnology / v.34, no.4, 2007 , pp. 375-380 More about this Journal
Abstract
This study was conducted to develop herbicide-resistance domestic maize plants by introducing the CP4 5-enol-pyruvylshikimate-3-phosphate synthase (CP4 EPSPS) gene using Agrobacterium tumefaciens-mediated immature embryo transformation. Immature embryos of five genotypes (HW1, KL103, HW3, HW4, HW7) were co-cultivated with strains Agrobacterium tumefaciens (strain C58C1) containing the binary vector (pCAMBIA2300) carrying Ubiquitin promoter-CP4 EPSPS gene and Cauliflower mosaic virus 35S (CaMV35S) promoter-nptll gene conferring resistance to paromomycin as a selective agent. The presence and expression of CP4 EPSPS transgene were confirmed by PCR, RT-PCR and Northern blot analysis, respectively. Also, the resistance to glyphosate in the transgenic maize ($T_1$) was analyzed by shikimate accumulation assay. The frequency (%) of paromomycin-resistance callus was 0.37, 0.03, 2.20, 2.37, and 0.81% in pure lines HW1, KL103, HW3, HW4 and HW7, respectively. EPSP transgene sequences were amplified in putative transgenic plants that regenerated from paromomycin-resistance calli of two inbred lines (HW3, HW4). Of them, RT-PCR and Northern blot analyses revealed that the transgene was only expressed in two transgenic events (M266, M104) of HW4 inbred line, and a mild glyphosate resistance of transgenic event (M266) was confirmed by the lower shikimate accumulation in leaf segments. These results demonstrate that transgenic maize with herbicide-resistance traits in Korean genotype can be genetically obtained.
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