广西医科大学学报2018，Vol.35Issue(4)：436-440,5.DOI:10.16190/j.cnki.45-1211/r.2018.04.004
TREM2基因过表达慢病毒载体的构建及其表达的研究
Construction and expression of lentiviral vector of TREM2 gene over-expression
摘要
Abstract
Objective:To construct lentiviral vector of a mouse triggering receptor expressed on myeloid cells-2,TREM2) gene over-expression,and examine the expression level of TREM2 in vitro.Methods:The TREM2 gene fragment was amplified by PCR,and the amplified product was ligated to the linearized GV358 vector.After identifying with PCR and DNA sequencing,the PCR product was co-transfected with the packaging plasmid into 293T cells and the virus titer was determined by drug screening method.293T cells were infected with recombinant lentivirus.GFP expression was observed and the expression level of TREM2 was detected by real-time quantitative PCR.Results:The TREM2 gene fragment was successfully obtained by PCR and ligated to the GV358 vector.TREM2-GV358 was confirmed to carry the correct TR(E)M2 gene by PCR and DNA sequencing.The virus was packaged in 293T cells and the virus titer was 2.0 ×108TU/mL.A large number of significant fluorescent cells infected by recombinant lentivirus were observed.Real-time quantitative PCR showed that the expression level of TREM2 was significantly increased(P＜0.05),when 293T cells was infected by lentivirus of TREM2 over-expression.Conclusion:The lentiviral vector over-expressing TREM2 gene was successfully constructed,which provides an experimental foundation for further studies on the functions of TREM2.关键词
TREM2/慢病毒/载体构建/过表达/293T细胞Key words
TREM2/lentivirus/vector construction/over-expression/293T cells分类
医药卫生引用本文复制引用
冉江霞,石胜良,王成志..TREM2基因过表达慢病毒载体的构建及其表达的研究[J].广西医科大学学报,2018,35(4):436-440,5.基金项目
国家自然科学基金资助项目(No.81460183) （No.81460183）
