遗传性长QT综合征HERG基因真核表达载体的构建和在HEK293细胞中的表达

目的构建HERG基因的真核表达载体，并在人胚胎肾细胞（HEK293）中进行表达。方法先将pGH19-HERG通过限制性酶SacⅠ和EcoR Ⅰ酶切得到HERG cDNA，将pIRES2-EGFP用SacⅠ和EcoRⅠ进行双酶切，把HERGcDNA定向克隆到pIRES2-EGFP中，即构建了HERG基因的真核表达载体pIRES2-EGFP—HERG。然后利用电穿孔法将pIRES2-EGFP-HERG转染HEK293细胞。结果在卵母细胞异源表达载体pGH19-HERG基础上，获得了真核表达载体pIRES2-EGFP—HERG，并在HEK293细胞中成功进行了表达。结论在HERG基因卵母细胞异源表达载体的基础上，构建了HERG基因的真核表达载体piRES2-EGFP-HERG，利用电穿孔法将其转染至HEK293细胞中并成功地进行了表达，为下一步进行膜片钳的研究奠定基础。

Construction of eukaryotic expression vector of LQTS HERG gene and its expression in HEK293 cell

Objective] To construct the eukaryotic expression vector of the HERG gene and express it in HEK293 cells. Methods] HERG cDNA was obtained from pGH19-HERG by restriction endonucleases Sac I and EcoR I digestion and was inserted into the same restriction site of pIRES2-EGFP in the same way, thus pIRES2- EGFP-HERG was constructed. The recombinant plasmids pIRES2-EGFP-HERG were identified by sequencing, pIRES2-EGFP-HERG was transfected into HEK293 cells by electroporation. Results] The eukaryotic expression vector pIRES2-EGFP-HERG was constructed correctly and expressed in HEK293 cells. Conclusions] The recombined pIRES2-EGFP-HERG eukaryotic expression vector can be expressed in HEK293 cells. These results make a foundation for the further functional study of HERG gene.