GFP基因在新疆小拟南芥和拟南芥中的表达分析  

Expression Analysis of GFP Gene in Olimarabidopsis pumila and Arabidopsis thaliana

作　　者：院海英[1] 吴祥辉[1] 东锐[1] 崔百明[1] 黄先忠[1] 

机构地区：[1]石河子大学生命科学学院,农业生物技术重点实验室,新疆石河子832003

出　　处：《西北植物学报》2012年第5期881-885,共5页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金项目(31060149);石河子大学高层次人才启动项目(RCZX200902)

摘　　要：以质粒pMCB30为模板,扩增GFP基因,连接到载体pCMBIA2300-35S-OCS上,构建过量表达载体p35S:GFP,将其转入农杆菌GV3101.通过农杆菌介导法将p35S:GFP载体分别转入新疆特色植物小拟南芥和拟南芥中.T0代经含有卡那霉素的1/2MS培养基筛选,获得了T1代转基因小拟南芥2株,T1代转基因拟南芥9株.通过激光共聚焦显微镜观察,在转基因小拟南芥和拟南芥的根尖细胞中均可检测到GFP绿色荧光蛋白;对转基因植株进行PCR扩增,均可检测到GFP基因,表明GFP基因已成功转入小拟南芥和拟南芥中.该研究建立了小拟南芥的遗传转化体系,为进一步利用GFP基因和进一步研究小拟南芥的功能基因奠定基础.GFP gene was amplified by PCR using the plasmid of pMCB30 as template, and the corrected GFP fragment was Iigated into the plant expression vector of pCMBIA2300-35S-OCS. The p35S：GFP vec tor was transformed into Agrobacterium GV3101 ,then p35S：GFP transferred into Olimarabidopsis pumila and Arabidopsis thaliana through Agrobacteriurn-mediated method. 2 independent plants of T, genera- tion of transgenic O. purnila and 9 independent plants of T1 generation of transgenic A. thaliana were screened on 1/2 MS medium containing kanamycin. GFP protein can be detected in the root tip cells of transgenic O. purnila and A. thaliana by confoeal laser microscopy. GFP gene was confirmed by PCR am= plification using DNA of transgenic plants,indicated that GFP gene had been successfully transformed into transgenic plants. The study layed foundations for further using of GFP gene and set up the genetic trans- formation system of O. pumila ,which will benefit a lot in further study of the functional genes of O. pumila.

关 键 词：GFP蛋白 拟南芥 小拟南芥 转基因 

分 类 号：Q786[生物学—分子生物学]