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学科主题基础医学
Establishment of HEK293 cell line expressing green fluorescent protein-aquaporin-1 to determine osmotic water permeability
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
2005-07-01
DOI10.1016/j.ab.2005.03.033
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
引用统计
被引频次:8[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/65563
专题北京大学基础医学院_药理学系
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
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GB/T 7714
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.
APA Gao, JW,Yu, HM,Song, QL,&Li, XJ.(2005).Establishment of HEK293 cell line expressing green fluorescent protein-aquaporin-1 to determine osmotic water permeability.ANALYTICAL BIOCHEMISTRY,342(1),53-58.
MLA Gao, JW,et al."Establishment of HEK293 cell line expressing green fluorescent protein-aquaporin-1 to determine osmotic water permeability".ANALYTICAL BIOCHEMISTRY 342.1(2005):53-58.
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