光诱导技术在Sf9昆虫细胞基因表达调控中的应用  

Application of Light-induction Technology to the Regulation of Gene Expression in Sf9 insect cells

作　　者：韩增鹏 蔡泽蕲 阮士龙 邵林 樊贵安 何晓斌 吴阳 徐富强 HAN Zeng-peng;CAI Ze-qi;RUAN Shi-long;SHAO Lin;FAN Gui-an;HE Xiao-bin;WU Yang;XU Fu-qiang(State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Key Laboratory of Magnetic Resonance in Biological Systems, Wuhan Center for Magnetic Resonance, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan,Hubei,430071,China;University of Chinese Academy of Sciences,Beijing,100049,China;Daye Public Inspection and Test Centre, Daye, Hube, 435100, China.)

机构地区：[1]中国科学院精密测量科学与技术创新研究院波谱与原子分子物理国家重点实验室,生物磁共振分析重点实验室,武汉磁共振中心,湖北武汉430071 [2]中国科学院大学,北京100049 [3]大冶市公共检验检测中心,湖北大冶435100

出　　处：《动物医学进展》2020年第12期50-55,共6页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31500868);中国科学院战略性先导科技专项项目(B类)(XDB32030200)。

摘　　要：光诱导技术是一种新兴的基因表达调控技术,目前已应用于原核细胞、哺乳动物和鱼类细胞。为探索光诱导技术在昆虫细胞基因表达系统中的调控功能,采用具有代表性的C120-EL222光诱导系统,在昆虫细胞表达质粒pMIB/V5-His的基础上,构建了集光诱导外源基因表达框与光激活转录因子于一体的昆虫细胞光诱导表达载体pMIB-C120-GOI-GFP-EL222质粒。将上述质粒转染昆虫Sf9细胞,通过检测光诱导产生的红色荧光蛋白(mCherry)或海参荧光素酶(hRluc)报告基因的表达,对光诱导元件的功能进行了定性和定量分析。结果表明,C120-EL222光诱导系统在昆虫Sf9细胞中具有较高的效能和严谨性,为光诱导表达技术在昆虫细胞基因表达调控中的深入研究和应用奠定了基础。Light-induction is a new technique of gene expression regulation.However,its application is limited to prokaryotic cells and mammalian and fish cells.In order to explore the functional effectiveness of light induction technology in insect cell lines,a representative C120-EL222 light induction system was adopted in this study.On the basis of insect cell expression plasmid pMIB/V5-His,the pMIB-C120-GOI-GFP-EL222 plasmid,which integrates the expression frame of exogenous gene induced by light and the transcription factor activated by light,was constructed.The above plasmids were transfected into insect Sf9 cells,and the expression of the light-induced red fluorescent protein(mCherry)or humanized Renilla luciferase(hRluc)reporter gene was detected after light induction,and the function of the light-induced elements was detected qualitatively and quantitatively.The experimental results showed that the C120-EL222 light induction system had high efficiency and preciseness in insect Sf9 cells.This study expands the application scope of the light-induced expression technology and lays a foundation for further research and application of the light-induced expression technology in insect cells.

关 键 词：光诱导技术 昆虫细胞系 基因表达调控 报告基因 

分 类 号：Q786[生物学—分子生物学] S852.74[农业科学—基础兽医学]