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KMID : 0613820150250020158
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Journal of Life Science
2015 Volume.25 No. 2 p.158 ~ p.163
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Optimized Germination Conditions and Human p53 Expression of Rice Embryo
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Pih Kyung-Tae
Choi Ju-Youn
Kim Keun-Cheol
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Abstract
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Rice embryo is more abundant than endosperms in nutrients such as proteins, lipids, and vitamin B1. In this study, we constructed p53 plasmid that could be expressed in a plant system, and investigated optimal germination conditions in a variety of media. For construction of p53 plasmid, we performed p53 amplification from pCDNA-p53, subcloned to TA cloning vector, and then reconstructed into pGEM-CaMV plant expression vector. On the other hand, we prepared a variety of imbibition buffers and complete media for efficient germination of the rice embryo. Imbibition buffers prepared with different concentrations of salt or detergent showed no significant effect on germination efficiency. We prepared further culture media, such as solid agar, liquid media, and paper towel to establish the optimal conditions. Rice embryo showed germination rates of more than 70% in the solid medium, more than 60% in the paper towel medium, but less than 25% in liquid media, although germination rate did not differ with varying concentrations of salt and sucrose in culture media. Under the optimal germination conditions, we introduced the p53 plasmid using imbibition method, and finally detected human p53 gene expression in the germinated rice embryo. This method might present a novel, practical approach for evaluating efficient gene expression utilizing imbibition method in rice embryo.
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KEYWORD
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DNA imbibition, expression, p53, rice embryo, solid medium
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