CAJ | 학술논문

　　目的：克隆人心房肌小电导钙激活钾通道亚型2（SK2），构建稳定表达SK通道的HEK293细胞系并研究其基本电生理学特性。方法：以人心房肌为标本，通过逆转录，重叠PCR法和定向克隆构建真核表达载体pIRES-hrGFP-SK2，采用脂质体法转染至HEK293细胞并筛选稳定表达的细胞系；使用全细胞和单通道膜片钳技术进行电流记录，研究其基本电生理学特性。结果：全细胞和单通道膜片钳均能在表达的HEK293细胞上记录到SK2通道电流，其电流具有明显的胞内钙离子依赖性。结论：成功构建稳定表达人心房肌SK2通道的HEK293细胞系，为以后研究通道功能活动奠定了基础。
　　목적：극륭인심방기소전도개격활갑통도아형2（SK2），구건은정표체SK통도적HEK293세포계병연구기기본전생이학특성。방법：이인심방기위표본，통과역전록，중첩PCR법화정향극륭구건진핵표체재체pIRES-hrGFP-SK2，채용지질체법전염지HEK293세포병사선은정표체적세포계；사용전세포화단통도막편겸기술진행전류기록，연구기기본전생이학특성。결과：전세포화단통도막편겸균능재표체적HEK293세포상기록도SK2통도전류，기전류구유명현적포내개리자의뢰성。결론：성공구건은정표체인심방기SK2통도적HEK293세포계，위이후연구통도공능활동전정료기출。
Objective: To clone the small conductance calcium activated potassium channels type 2 (SK2)of human atrial tissue,construct stable HEK293 cells expressed SK2 and investigate the basic electrophysiological properties. Methods: The expression plasmid of pIRES-hrGFP-SK2 was constructed by reverse transcription, overlapping PCR and site-directed cloning. The plasmid was transfected by Lipofectamine 2000 and stable cell lines expressed SK2 were selected. The basic properties of SK2 were investigated by using patch clamp technique. Results: SK2 currents could be recorded in HEK293 cells expressed SK2 channels in whole-cell and single-channel configuration. SK2 currents were calcium dependenent. Conclusion: Stable HEK293 cells expressed SK2 are constructed successfully, which establishes the basis for further research about the regulation of SK2 channels.