郑州大学学报（医学版）2016，Vol.51Issue(5)：580-582,583,4.DOI:10.13705/j.issn.1671-6825.2016.05.006
SK2和 JPH2双基因重组真核表达载体的构建及在 HEK293细胞中的表达
Construction of an eukaryotic expression vector carried SK 2 and JPH2 genes and expressions of SK2 and JPH2 proteins in HEK293 cells
摘要
Abstract
Aim:To construct an eukaryotic expression vector carried SK2 and JPH2 genes and detect the expression of them in the transfected HEK293 cells.Methods:The full-length SK2 gene was obtained from a recombinant vector pC-MV6-entry/SK2 by PCR amplification, and cloned it into the eukaryotic expression vector pIRES -EGFP.Then JPH2 gene was inserted into pIRES-EGFP-SK2 vector and was detected by endonuclease digestion and sequencing .The recombinant pIRES-EGFP/SK2+JPH2 was transfected into HEK293 cells with a liposome infection protocol.The expressions of SK2 and JPH 2 protein were detected by Western blot.Results:Both endonuclease digestion and sequence analysis demonstrated that the inserted sequences of pIRES-EGFP/SK2+JPH2 were identical to those of the full-length of SK2 and JPH2 genes in GenBank.Moreover, both SK2 and JPH2 protein in HEK 293 cells transfected with pIRES-EGFP/SK2+JPH2 for 48 h were expressed successfully.Conclusion: The eukaryotic expression vector pIRESE-GFP/SK2+JPH2 as well as the ex-pressions of SK2 and JPH2 cell lines have been successfully constructed .关键词
SK2通道/JPH2蛋白/真核表达载体/HEK293Key words
SK2 channel/JPH2 protein/eukaryotic expression vector/HEK293 cell line分类
医药卫生引用本文复制引用
罗天霞,樊红琨,芮丹丹,孙佳翕,马小芳,章茜..SK2和 JPH2双基因重组真核表达载体的构建及在 HEK293细胞中的表达[J].郑州大学学报（医学版）,2016,51(5):580-582,583,4.基金项目
国家自然科学基金资助项目81270248；81570311 （）
