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学科主题: 基础医学
题名:
Establishment of HEK293 cell line expressing green fluorescent protein-aquaporin-1 to determine osmotic water permeability
作者: Gao, JW; Yu, HM; Song, QL; Li, XJ
关键词: osmotic water permeability ; fluorescence density ; transfected cell ; aquaporin-1 (AQP1)
刊名: ANALYTICAL BIOCHEMISTRY
发表日期: 2005-07-01
DOI: 10.1016/j.ab.2005.03.033
卷: 342, 期:1, 页:53-58
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]: XENOPUS OOCYTES ; CHANNELS ; PROTEIN ; LOCALIZATION ; KIDNEY ; CHIP28
英文摘要:

Aquaporin (AQP) is a kind of channel-forming membrane glycoprotein that mediates osmotic water transport. The present study aimed to establish a cell line stably transfected with AQP1 to measure osmotic water permeability. The recombinant plasmid was constructed by subcloning the full-length rat AQP1 cDNA into pEGFP-C3 vector, named pEGFP/AQP1. Human embryonic kidney 293 cells were transfected with pEGFP/AQP1 and selected by G418 to obtain a cell line stably expressing AQP1 tagged with green fluorescent protein. The expression level of AQPI in the stably transfected cell was detected by reverse transcription polymerase chain reaction and Western blot. The real-time change of fluorescence density, corresponding to cell swelling induced by hyposmotic solution, was recorded under confocal laser scanning microscope and used to assess osmotic water permeability. The typical AQP1 inhibitor, mercuric chloride, validated this osmotic water permeability assay. These results suggested that this transfected cell model could be conveniently used to determine osmotic water permeability. (c) 2005 Elsevier Inc. All rights reserved.

语种: 英语
WOS记录号: WOS:000230277100007
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/65563
Appears in Collections:基础医学院_药理学系_期刊论文

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作者单位: 1.Peking Univ, Hlth Sci Ctr, Dept Pharmacol, Beijing 100083, Peoples R China
2.Peking Univ, State Key Lab Nat & Biomimet Drugs, Beijing 100083, Peoples R China
3.Natl Res Inst Family Planning, Beijing 100083, Peoples R China

Recommended Citation:
Gao, JW,Yu, HM,Song, QL,et al. Establishment of HEK293 cell line expressing green fluorescent protein-aquaporin-1 to determine osmotic water permeability[J]. ANALYTICAL BIOCHEMISTRY,2005,342(1):53-58.
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