昆明医科大学学报2013，Vol.34Issue(7)：4-7,4.
用慢病毒转染技术实现Gnaq基因在SH-SY5Y细胞中持续稳定高表达
Persistant and Stable Over-Expression of Gnaq in SH-SY5Y Cell Line by Means of Lentivector Transfection Technique
摘要
Abstract
Objective To probe the feasibility of overexpression of Gnaq in SH-SY5Y cell line by means of lentivector transfection technique so as to accumulate the necessary basis for further studies of the roles and molecular mechanisms of Gnaq in brain aging and related diseases.Methods The CDS of Gnaq in NCBI website were searched and corresponding cloning primers were designed.The target gene which was obtained by PCR amplifying the eDNA of HepG2 cell was cloned into lentivector PLIG.Recombinant plasmid PLIG-Gnaq was transfected into competent cells and positive clones were selected.After being verified by means of sequence,lentivirus were generated to transfect SH-SY5Y cell.GFP was measured at 24 hours after transfection and the 5th subculture.Transcription level of Gnaq was detected at the 3rd and 5th subculture respectively.Results The efficient rate of transfetion of Gnaq gene into SH-SY5Y cell was about 100％.GFP expressed that almost all of the cells continuously went on to the 5th subculture.The transcription level of Gnaq was significantly enhanced both in the 3rd and 5th subcultures of PLIG-Gnaq-SH-SY5Y cells.Conclusion It is feasible to reach the persistent and stable overexpression of Gnaq in SH-SY5Y cell line by means of lentivector transfection technique.The PLIG-Gnaq-SH-SY5Y cells can be used as model cells for further studies of the roles and underlying mechanisms of Gnaq in neurons.关键词
慢病毒载体/Gnaq基因/SH-SY5Y细胞/过表达Key words
Lentivector/ Gnaq gene/ SH-SY5Y cell line/ Over-expression分类
生物科学引用本文复制引用
陈绍春,邹智荣,李国萍,叶频,范艳,李跃敏..用慢病毒转染技术实现Gnaq基因在SH-SY5Y细胞中持续稳定高表达[J].昆明医科大学学报,2013,34(7):4-7,4.基金项目
国家自然科学基金资助项目(81260195),云南省自然科学基金资助项目(2010ZC110) （81260195）
