| 稳定表达hHCN2基因 HEK293细胞系的建立 |
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| 作者姓名： | Zhao X Yang XJ Bai X Li HX Cheng XJ Jiang WP |
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| 作者单位： | 1. 苏州大学附属第一医院心内科,江苏,苏州,215006
2. 江苏血液病研究所,江苏,苏州,215006 |
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| 基金项目： | 苏州大学校科研和校改项目 |
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| 摘 要： | 目的：培育稳定表达hHCN2基因的细胞系,建立一种表达研究心肌离子通道的有效模型。方法：通过脂质体转染的方法,将重组pcDNA3-hHCN2真核表达载体导入人胚肾细胞（HEK293细胞）,以G418压力筛选转染细胞,并对其进行全细胞膜片钳记录。结果：经600μg／ml压力筛选后,获得抗性细胞克隆,并用全细胞膜片钳技术记录到克隆hHCN2通道编码电流。结论：本实验采用脂质体转染法成功地培育出G418抗性HEK293细胞。为进一步研究克隆离子通道结构和功能的关系奠定基础。
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| 关 键 词： | 细胞系 基因转染 膜片钳 |
| 文章编号： | 1000-6834（2006）02-0254-03 |
| 收稿时间： | 2005-07-08 |
| 修稿时间： | 2005-07-082006-01-04 |
Establishment of a HEK293 cell line stably expressing human HCN2 gene |
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| Zhao X,Yang XJ,Bai X,Li HX,Cheng XJ,Jiang WP.Establishment of a HEK293 cell line stably expressing human HCN2 gene[J].Chinese Journal of Applied Physiology,2006,22(2):254-256. |
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| Authors: | Zhao Xin Yang Xiang-Jun Bai Xia Li Hong-Xia Cheng Xu-Jie Jiang Wen-Ping |
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| Institution: | 1. Department Cardiovascular Research, The First Affiliated Hospital to Suzhou University, 2. Jiangsu Institute of Hematology, Suzhou 215006, China |
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| Abstract: | Aim: To create a model for studying ionic channels by means of the expressing human HCN2 and G418-resistant HEK293 cell lines established.Methods: pcDNA3-hHCN2 was transfected with Lipofectin2000 into HEK293 cell line.The transfected cells would be survived in the further culture medium containing G418 antibiotic as the hHCN2 gene could express a G418 resistant products.Whole-cell patch clamp investigated that hHCN2 gene was transfected into HEK293 cells.Results: The G418 resistant(600ug/ml) HEK293 cell line was established successfully and whole-cell patch clamp recorded ionic currents of transfected hHCN2.Conclusion: The G418 resistant HEK293 cell line was successfully established with transfection of plasmid pcDNA3-hHCN2 by Lipofectin,which might be useful for studying the relationship between the structure and function of cloned ionic channels. |
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| Keywords: | cell line gene transfection patch clamp |
| 本文献已被 CNKI 维普 万方数据 PubMed 等数据库收录！ |
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