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http://dx.doi.org/10.3839/jabc.2010.034

The Optimization for Functional Expression of Arabidopsis Thaliana AtPIP2-1 in Xenopus laevis Oocyte  

Kim, Hyun-Mi (Bio-crop Development Division, National Academy of Agricultural Science, Rural Development Administration)
Hwang, Hyun-Sik (Bio-crop Development Division, National Academy of Agricultural Science, Rural Development Administration)
Lee, Suk-Chan (Department of Genetic Engineering, Sungkyunkwan University)
Jo, Su-Hyun (Department of Physiology, Kangwon National University College of Medicine)
Kim, Beom-Gi (Bio-crop Development Division, National Academy of Agricultural Science, Rural Development Administration)
Publication Information
Journal of Applied Biological Chemistry / v.53, no.4, 2010 , pp. 189-194 More about this Journal
Abstract
We confirmed the hypo-osmotic shock strengths and duration, different type of vectors, and subcelluar localization to identify the optimum analysis condition of plant aquaporin activity in Xenopus ooctye using Arabidopsis thaliana AtPIP2-1 gene. Six minutes and 1/5ND buffer hypoosmotic shock treatment was the best condition to show the maximum swelling of Xenopus oocytes where AtPIP2-1 was expressed using pcDNA3.1 vector. AtPIP2-1 protein was expressed more efficiently in pGEMHE vector which has 5' and 3' UTR (untranslation region) of Xenopus ${\beta}$-GLOBIN gene in multiple cloning site than in pcDNA3.1 vector. Also green fluorescence of GFP fused to AtPIP2-1 was detected onto oocyte plasmamembrane where is the proper subcellular localization target of AtPIP2-1.
Keywords
aquaporin activity; AtPIP2-1; Xenopus oocyte;
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