생화학/분자생물학 : Xenopus oocyte에서 애기장대 AtPIP2-1 활성측정을 위한 발현 최적화 조건 규명

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 treatme...

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Bibliographic Details
Published in:Journal of applied biological chemistry 2010-12, Vol.53 (4), p.189
Main Authors: 김현미, Hyun Mi Kim, 황현식, Hyun Sik Hwang, 이석찬, Suk Chan Lee, 조수현, Su Hyun Jo, 김범기, Beom Gi Kim
Format: Article
Language:Korean
Subjects:
aquaporin activity
AtPIP2-1
Xenopus oocyte
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Summary: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 β-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.
ISSN:1976-0442