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作 者 ：院海英，吴祥辉，东 锐，崔百明，黄先忠*

期 刊 ：西北植物学报 2012年 5期 页码：881~885

关键词：GFP蛋白；拟南芥；小拟南芥；转基因；

Keywords:GFP, Arabidopsis thaliana, Olimarabidopsis pumila, transgene,

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1/2 MS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。

Abstract:GFP gene was amplified by PCR using the plasmid of pMCB30 as template,and the corrected GFP fragment was ligated into the plant expression vector of pCMBIA2300-35S-OCS.The p35S:GFP vector was transformed into Agrobacterium GV3101,then p35S:GFP transferred into Olimarabidopsis pumila and Arabidopsis thaliana through Agrobacterium-mediated method.2 independent plants of T₁ generation of transgenic O.pumila 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.pumila and A.thaliana by confocal laser microscopy.GFP gene was confirmed by PCR amplification 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 transformation system of O.pumila,which will benefit a lot in further study of the functional genes of O.pumila.

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