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水通道蛋白-9表达载体的构建及视神经脊髓炎患者血清中抗体的检测

高聪 郑扬波 单福兰 范永祥 吴林展 钟容 龙友明

广东医学Issue(6):821-824,4.
广东医学Issue(6):821-824,4.

水通道蛋白-9表达载体的构建及视神经脊髓炎患者血清中抗体的检测

Construction of aquaporin-9 expressing vector and detection of AQP-9 antibody in patients with neuromyelitis optica

高聪 1郑扬波 1单福兰 1范永祥 1吴林展 1钟容 1龙友明1

作者信息

  • 1. 广州医科大学附属第二医院神经内科 广州医科大学附属第二医院神经科学研究所 广州510260
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摘要

Abstract

Objective To construct a eukaryotic aquaporin -9 (AQP9) expressing vector for AQP9 antibody de-tection in patients with neuromyelitis optica (NMO).Methods The recombinant vector was constructed by subcloning human AQP9 cDNA into pcDNA3.1+vector, with which human embryonic kidney (HEK) 293T cells was transfected to establish an AQP9 protein expressing cell line .The cell line expression AQP 9 was confirmed by reverse transcription poly-merase chain reaction ( RT-PCR) and Western blot analysis .Serum antibody was tested with cell -based assay by indi-rect immunofluorescence assay .Results The relative expression level of AQP 9 mRNA was significantly higher than the empty vector, and Western blotting analysis showed the expression content at 32 kD bands was significantly higher than the empty vector and 293T cells.The cell line expressing AQP9 was successfully constructed and confirmed by reverse tran-scription polymerase chain reaction ( RT-PCR) and Western blot analysis .The serum antibody was positive in all 4 pa-tients with NMO and negative in one with cerebral infarction and one normal adult .Conclusion The transfected cell mod-el is simple and advantageous in AQP 9 serum antibody detection for research on NMO .

关键词

水通道蛋白-9/抗体/视神经脊髓炎

Key words

Aquaporin-9/neuromyelitis optica/HEK 293T/antibody

引用本文复制引用

高聪,郑扬波,单福兰,范永祥,吴林展,钟容,龙友明..水通道蛋白-9表达载体的构建及视神经脊髓炎患者血清中抗体的检测[J].广东医学,2015,(6):821-824,4.

基金项目

广东省自然科学基金资助项目 ()

广东医学

OA北大核心CSTPCD

1001-9448

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