泸州医学院学报2016，Vol.39Issue(6)：527-530,4.DOI:10.3969/j.issn.1000-2669.2016.06.009
Flag和GFP双标记的SK2通道表达质粒的构建尧鉴定和序列分析
Construction of SK2 channel expression plasmid with double labeling of Flag and GFP
摘要
Abstract
Objective: This study aimed to construct a SK2 channel plasmid with two tags using overlapping PCR to provide the easiness to track the transportation of the channel protein. Methods: Based on human SK2 channel expression plasmid pIRES-SK2, the full length of SK2 cDNA with a Flag insertion was amplified using overlapping PCR method, which was then subcloned into pEGFP-N3 vector. The double-tagged expression plasmid, pEGFP-N3-Flag-SK2 (Flag-SK2-GFP) was confirmed by DNA sequencing. Results: A Flag tag was inserted in-between the S1-S2 extracellular loops of SK2 channel whose C-terminus was fused in-frame to GFP. The constructed plasmid was further confirmed by DNA sequencing. Conclusion: The expression plasmid Flag-SK2-GFP was constructed successfully with overlapping PCR.关键词
小电导钙激活钾通道/重叠PCR/基因工程Key words
Small conductance calcium activated potassium channels/Overlapping PCR/Gene cloning分类
药学引用本文复制引用
黄文俊,李涛,范学慧,余奕言,杨艳,曾晓荣,谭晓秋..Flag和GFP双标记的SK2通道表达质粒的构建尧鉴定和序列分析[J].泸州医学院学报,2016,39(6):527-530,4.基金项目
国家自然科学基金资助项目（NO：31300948；81670310），四川省科技厅支撑计划（2011FZ0106），泸州市-四川医科大学联合资助项目 （）
