中南医学科学杂志2017，Vol.45Issue(2)：132-135,4.DOI:10.15972/j.cnki.43-1509/r.2017.02.006
SCN3A基因突变型表达载体的构建及HEK 293细胞中的表达
Construction of SCN3A gene mutant expression vector andits expression in HEK 293 cells
摘要
Abstract
Objective To construct the mutant N302S (c.905A＞G) of plasmid pCMV6-XL4-SCN3A containing the SCN3A gene mRNA fragment in vitro,and provide experimental basis for further functional studies.Methods The primers with mutation sites were designed and the mutant plasmids were constructed by site-directed mutagenesis using wild-type plasmids as templates.The recombinant plasmid was transiently transfected into HEK293 cells,and the expression of the target gene was verified by RT-PCR after 24 hours.Results Recombinant plasmid digestion and sequencing confirmed that the mutation was successful.The expression of the target gene was detected by RT-PCR after transfection of recombinant SCN3A eukaryotic expression plasmid into HEK293 cell line.Conclusion The SCN3A gene mutant eukaryotic expression vector pCMV6-XL4-SCN3A-N302S was successfully constructed and expressed in HEK293 cells at the gene level.关键词
SCN3A/质粒/表达/RT-PCRKey words
SCN3A/plasmid/expression/RT-PCR分类
医药卫生引用本文复制引用
陈勇军,邱国真,汤斌,刘稀金,李欣,何妍妍..SCN3A基因突变型表达载体的构建及HEK 293细胞中的表达[J].中南医学科学杂志,2017,45(2):132-135,4.基金项目
湖南省教育厅立项课题资助(15C1206)及衡阳市科技局课题资助(2015KJ59). （15C1206）
