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《Journal of Medical Postgraduates》 2007-11
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Cloning of KCNE1 gene in long QT syndrome and construction of its eukaryotic expression vector

LI Wei1,YANG Jun-guo2,DU Rong2,XU Qiu-mei1,GUAN Si-ming1,KE Qin-mei1,WANG Bin1(1.Department of Geriatrics;2.Institute of Cardiology,Xiehe Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,Hubei,China)  
Objective: To construct the eurokaryotic expression vector of KCNE1 gene and express recombinant KCNE1 in HEK293 cells.Methods:Human KCNE1 gene fragment was amplified from human placenta total RNA by RT-PCR and cloned into the vector of pCR2.1 TOPO by means of T-A cloning.KCNE1 cDNA was obtained from pCR2.1-KCNE1 by restriction enzyme digestion and inserted into the same restriction site of pEGFP-N1.Thus pEGFP-N1-KCNE1 was constructed and transfected into HEK293 cells with Effectene transfection reagent.Results:The eukaryotic expression vector pEGFP-N1-KCNE1 was successfully constructed by gene cloning and recombinant method and expressed in HEK293 cells.Conclusion:The cloning of KCNE1 gene and the construction and expression of its eukaryotic expression vector may shed some light on further functional study of KCNE1 gene.
【Key Words】： Long QT syndrome KCNE1 gene Clone Eukaryotic expression vector
【Fund】： 国家自然科学基金资助项目(批准号:30170377);; 国家科技攻关计划863项目资助(批准号:2002BA711A07)
【CateGory Index】： R541.7
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