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http://dx.doi.org/10.5010/JPB.2016.43.3.341

Production of transgenic cucumber expressing phytoene synthase-2A carotene desaturase gene  

Jang, Hyun A (Department of Biology, Chungnam National Univ.)
Utomo, Setyo Dwi (Department of Agronomy and Horticulture, Faculty of Agriculture University of Lampung)
Kwon, Suk Yoon (Korea Research Institute of Bioscience and Biotechnology)
Ha, Sun-Hwa (Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University)
Xing-guo, Ye (Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement)
Choi, Pil Son (Department of Medicinal Plant Resources, Nambu University)
Publication Information
Journal of Plant Biotechnology / v.43, no.3, 2016 , pp. 341-346 More about this Journal
Abstract
The objectives of this study were to 1) evaluate the efficiency of the protocol of Agrobacterium-mediated transformation of cucumber to introduce phytoene synthase-2a carotene desaturase (PAC genes); 2) demonstrate the integration of PAC genes into the genome of putative transgenic cucumber based on growth on selection medium, PCR and Southern analysis; 3) evaluate the expression of PAC genes in transgenic cucumber based on the analysis of RT-PCR and Northern blot hybridization. Out of 5,945 cotyledonary-node explants inoculated with Agrobacterium, 65 (1.1%) explants produced 238 shoots. Integration of PAC genes into the genome of the cucumber was demonstrated based on the analysis of gDNA-PCR, 21 out of the 238 plants regenerated; while 6 plants proved positive for Southern blot hybridization. Transgene expression was demonstrated based on analysis of RT-PCR, 6 plants proved positive out of the 6 plants analyzed; while 4 plants out of 6 proved positive during Northern blot hybridization. This study successfully demonstrated the production of transgenic cucumber, integration, and expression of the PAC gene in cucumber.
Keywords
${\beta}$-carotene; Cucumis sativus; cotyledonary-node explant; transformation efficiency;
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Times Cited By KSCI : 6  (Citation Analysis)
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