芪合酶基因Fm-STS在何首乌毛状根中的过量表达分析  

Over-expression of Stilbene Synthase Gene FM-STS in Hairy Roots of Polygonum Multiflorum

作　　者：朱宽鹏[1] 生书晶[1] 赵炜[1] 陆娣[1] 夏晚霞[1] 赵树进[1] 

机构地区：[1]广州军区广州总医院,广东广州510010

出　　处：《现代生物医学进展》2013年第5期832-836,979,共6页Progress in Modern Biomedicine

基　　金：广东省自然科学基金项目(10151001002000012);广东省科技计划(00697862100303016)

摘　　要：目的:通过过量表达探究在何首乌中得到的芪合酶基因Fm-STS的功能。方法:由含CaMV 35S启动子驱动以及荧光标记蛋白(Green fluorescent protein,GFP)基因的植物转基因基础表达质粒pBIN-35S-GFP构建过量表达质粒pBIN-35S-STS-GFP(阳性),并同空白表达质粒pBIN-35S-GFP(空白)均导入野生型发根农杆菌ATCC15834中,转化何首乌外植体(无菌苗叶片),诱导生成毛状根并培养,对毛状根进行高效液相色谱分析芪合物二苯乙烯苷含量变化以及实时荧光定量检测基因Fm-STS表达差异。结果:在过表达组和空白组中毛状根中发根农杆菌Ri质粒中的rolB基因和外源基因GFP均有表达,芪合物二苯乙烯苷含量依次为4.67 mg/g和2.18 mg/g(干重),在mRNA水平上检测基因Fm-STS表达量:过表达组是空白组的2.41倍。结论:基因FM-STS是何首乌中芪合物二苯乙烯苷生物合成过程中的酶基因,过量表达在基因功能研究中有良好的应用。Objective： Used over-expression of the gene FM-STS that come from Polygonum Multiflorum to research its function.Methods： The over-expression vector pBIN-35S-STS-GFP was constructed by the blank plasmid pBIN-35S-GFP and the gene FM-STS sequence.Sent the over-expression plasmid and the blank plasmid into wild-type Agrobacterium rhizogenes ATCC15834,then they induced hairy roots and cultured it.In the end we analyzed hairy roots by HPLC to determine the component stilbene glucoside and Real-time PCR to analysis expression of the gene FM-STS.Results： The PCR results showed that the gene rolB and GFP are both expressed in hairy roots.Finally,the content of Stilbene glucoside in blank group is 2.18mg/g and in the over-expression group is 4.67mg/g,At the mRNA level to detect gene Fm-STS expression level,in the over-expression group is the highest,it＇s 2.41 times in the blank group.Conclusion： The gene FM-STS is a enzyme gene in synthesis of stilbene glucoside,the method over-expression is good use in research of gene＇ s function.

关 键 词：过量表达 何首乌 毛状根 荧光定量PCR 高效液相色谱法 

分 类 号：S567.239[农业科学—中草药栽培] Q75[农业科学—作物学]