Cloning of KCNE1 gene in long QT syndrome and construction of its eukaryotic expression vector / 医学研究生学报

ABSTRACT

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.