| 摘要: |
| 为研究VR1（Vanilloid receptor subtype1）受体的功能，建立VR1稳定真核表达细胞系。［方法］ 采用 RT-PCR的方法从大鼠脊髓克隆VR1 cDNA，并构建VR1表达载体。将VR1质粒通过电穿孔方法转染HEK293细胞， 经G418筛选获得稳定表达的细胞克隆。采用RT-PCR、Western blotting、免疫荧光、细胞Ca2+浓度测定等方法检测VR1受体的表达。［结果］筛选的阳性细胞克隆含有VR1的mRNA及蛋白质，细胞Ca2+浓度的改变提示VR1受体的功能性表达。［结论］建立了稳定表达VR1受体的真核表达细胞系 |
| 关键词: VR1 HEK293细胞 细胞系 |
| DOI：10.11724/jdmu.2002.04.01 |
| 分类号:R962 |
| 基金项目: |
|
| Characterization of Vanilloid receptor subtype1 (VR1) stably expressed in HEK293 cells |
|
YAO Ji-hong1, KANAME Onozawa2, KAZUHIRO Kohama2
|
|
1.Department of Pharmacology, Dalian Medical University, Dalian, 116027 China;2.Department of Pharmacology, Gunma University, Japan
|
| Abstract: |
| In order to study the mechanism of Vanilloid receptor subtype1 (VR1), VR1 gene was expressed in cultured HEK293 cells. ［Method］ VR1 cDNA was cloned from rat dorsal root ganglion cells by an RT-PCR and a VR1-expressing plasmid was constructed. The plasmid was introduced into HEK293 cells. ［Result］ The stable cell lines expressing VR1 were confirmed by RT-PCR, Western blotting, immunofluorescence and calcium influx assay. ［Conclusion］ The VR1 is functional expressed in HEK293 cells; |
| Key words: Vanilloid receptor subtype1 (VR1) HEK293 cells |