| 阴离子交换蛋白1在HEK293t细胞中的高效表达与鉴定 |
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| 引用本文： | 席玉慧,白淑芝,傅国辉.阴离子交换蛋白1在HEK293t细胞中的高效表达与鉴定[J].哈尔滨医科大学学报,2005,39(2):107-110. |
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| 作者姓名： | 席玉慧 白淑芝 傅国辉 |
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| 作者单位： | 哈尔滨医科大学,病理生理学教研室,黑龙江,哈尔滨,150086;哈尔滨医科大学,病理生理学教研室,黑龙江,哈尔滨,150086;哈尔滨医科大学,病理生理学教研室,黑龙江,哈尔滨,150086 |
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| 基金项目： | 国家自然科学基金资助项目 (3 0 2 3 0 160 ) |
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| 摘 要： | 目的在人胚肾HEK293t(293t)细胞高效表达具有阴离子转运活性的阴离子交换蛋白1(AE1)。方法用传统磷酸钙转染法将质粒pRBG4-AE1转入293t细胞，免疫荧光及Western blotting验证蛋白表达及正常的细胞膜定位后，应用6-甲氧基-N-(3-磺丙基)喹啉铵(SPQ)观察表达的AE1是否具有阴离子转运活性。结果在293t细胞中，显示AE1高效表达。转染pRBG4-AE1后293t细胞相对于正常以及转染空载体对照组的阴离子交换功能明显增强。结论利用传统磷酸钙转染法，在293t可高效表达具有正常跨膜拓扑结构及阴离子转运活性的AE1，为进一步研究其相关功能奠定了基础。
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| 关 键 词： | 阴离子交换蛋白1 磷酸钙转染 蛋白表达 6-甲氧基-N-(3-磺丙基)喹啉铵 |
| 文章编号： | 1000-1905(2005)02-0107-03 |
| 修稿时间： | 2004年11月19 |
Expression and identification of anion exchanger 1 in HEK293t cells |
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| XI Yu-hui,BAI Shu-zhi,FU Guo-hui.Expression and identification of anion exchanger 1 in HEK293t cells[J].Journal of Harbin Medical University,2005,39(2):107-110. |
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| Authors: | XI Yu-hui BAI Shu-zhi FU Guo-hui |
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| Abstract: | Objective To express the functional anion exchanger 1(AE1) in HEK293t (293t) cells.Methods The cDNA encoding full length AE1 was transfected into 293t cells with calcium phosphate.Expression of AE1 in 293t cells was detected by western-blotting and fluorescence microscopy.6-methoxy-N-(3-sulfopropyl)quinolinium(SPQ)was used to detect the anion exchange activity of AE1.Results AE1 was efficiently expressed in the transfected cells and the anion exchange activity of these cells was obviously increased compared with the activity of the control.Conclusion AE1 could be abundantly expressed on the membrane of 293t cells with calcium phosphate and has normal transmembrane-topology and anion exchange activity.A useful cell model is provided for the further functional investigation of AE1. |
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| Keywords: | anion exchanger 1 calcium phosphate protein expression 6-methoxy-N-(3-sulfopropyl) quinolinium |
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