| 间隙连接蛋白26-EGFP载体的构建与表达 |
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| 引用本文： | 王红英,郑维国,陈必良,王德堂.间隙连接蛋白26-EGFP载体的构建与表达[J].医学研究生学报,2004,17(10):872-873,876,F002. |
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| 作者姓名： | 王红英 郑维国 陈必良 王德堂 |
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| 作者单位： | 第四军医大学西京医院妇产科,陕西西安,710032 |
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| 摘 要： | 目的：构建间隙连接蛋白(connexin，Cx)26-EGFP真核表达载体，观察其在细胞内的表达和功能情况。方法：经RT-PCR获得Cx26基因全长片段，重组至有增强绿色荧光蛋白标记的真核表达载体pEGFP-N2中，经脂质体转染至宫颈癌Hela细胞中，通过激光共聚焦显微镜、免疫细胞化学方法、蛋白印迹法(Western blot)检测蛋白表达，应用荧光染料(Lucifer Yellow)细胞划痕法检测细胞间间隙连接通信状态。结果：转染细胞有增强绿色荧光蛋白及Cx26的表达，细胞间间隙连接通讯恢复。结论：融合表达增强绿色荧光蛋白后，不影响间隙连接蛋白的功能，在间隙连接蛋白的研究中，pEGFP-N载体可以作为一个优选载体。
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| 关 键 词： | 间隙连接蛋白 绿色荧光蛋白 转染 |
| 文章编号： | 1008-8199(2004)10-0872-02 |
Creation and expression of a connexin 26-enhance green fluorescent protein chimera in Hela cells |
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| WANG Hong-ying,ZHENG Wei-guo,CHEN Bi-liang,WANG De-tang.Creation and expression of a connexin 26-enhance green fluorescent protein chimera in Hela cells[J].Bulletin of Medical Postgraduate,2004,17(10):872-873,876,F002. |
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| Authors: | WANG Hong-ying ZHENG Wei-guo CHEN Bi-liang WANG De-tang |
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| Abstract: | Objective:To investigate the expression and function of connexin26-EGFP gene in human cervical carcinoma cell line Hela. Methods:Connexin26-EGFP gene was created and transfected it into Hela cells. Laser scanning confocal microscope, immunohistochemical staining, and Western blot were used to detect the expression of Cx26-EGFP protein. Lucifer Yellow dye transfer was used to evaluate the function of the protein. Results:After being transfected, the cell express Cx26-EGFP protein. The result of dye transfer studies showed that Cx26-EGFP forming functional gap junction channels. Conclusion:Connexin-Enhance Green Fluorescent Protein Chimera formed functional gap junction channels, The pEGFP-N is an excellent vector in the study of gap junction. |
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| Keywords: | Connexin GFP Transfect |
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