mCLCA3增强型绿色荧光蛋白真核表达系统的构建及功能分析

Construction of eukaryotic expression system mCLCA3-enhanced green fluorescent protein and function analysis of mCLCA3 protein

目的建立mCLCA3真核细胞表达模型,并对其氯离子通道功能进行分析,为分析其分子本质提供新的理论依据。方法采用RT-PCR、脂质体转染法构建mCLCA3增强绿色荧光型真核细胞表达模型,通过免疫荧光对mCLCA3真核表达情况进行分析,利用FluoStar Optima荧光仪对mCLCA3的氯离子通道功能进行测定分析。结果构建真核表达模型能够稳定高表达mCLCA3及增强型绿色荧光蛋白,适用于离子通道功能分析。结论成功构建了mCLCA3增强绿色荧光型真核细胞表达模型,并通过功能分析证明mCLCA3可能不是氯离子通道。

Objective To eonstruet eukaryotie expression system mCLCA3-enhaneed green fluoreseent protein and analysis the ehloride ehannel funetion of mCLCA3 protein. Method Eukaryotic expression system mCLCA3-enhaneed green fluorescent protein was constructed by RT-PCR and liposome-transfeetion. The expression of mCLCA3 was determined by immune-florescence ; FluoStar Optima was used to detect the chloride channel function of mCLCA3. Results Eukaryotie expression system of mCLCA3-enhaneed green fluoreseent protein could be used to analysis the ehloride ehannel funetion of mCLCA3 protein. Conclusion Eukaryotie expression system of mCLCA3-enhaneed green fluorescent protein was construeted and the mCLCA3 protein may not be a chloride channel function.