| 番茄EBF2基因的克隆、亚细胞定位与遗传转化 |
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| 引用本文： | 吴玉,杨迎伍,邓伟,李正国.番茄EBF2基因的克隆、亚细胞定位与遗传转化[J].核农学报,2010,24(3):490-494. |
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| 作者姓名： | 吴玉 杨迎伍 邓伟 李正国 |
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| 作者单位： | 重庆大学生物工程学院基因工程研究中心,重庆,400030;重庆市高校功能基因与调控新技术重点实验室,重庆,400030 |
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| 摘 要： | 为研究番茄EBF2基因的功能,利用RT-PCR技术从番茄果实cDNA中扩增了全长EBF2基因,将其与绿色荧光蛋白(GFP)构建成融合表达载体,在烟草原生质体中进行瞬时表达。结果表明,该基因定位于细胞核内。将该基因定向克隆到植物表达载体pCambia1301,构建成由组成型启动子CaMV35S启动的植物表达载体pCambia1301-EBF2,通过农杆菌介导方法转化MicroTom番茄。经PCR及GUS组织染色检测,外源基因已整合到番茄基因组中。
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| 关 键 词： | EBF2基因 亚细胞定位 载体构建 遗传转化 番茄 |
CLONING, SUBCELLULAR LOCALIZATION AND TRANSFORMATION OF EBF2 GENE IN TOMATO |
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| WU Yu,YANG Ying-wu,DENG Wei,LI Zheng-guo.CLONING, SUBCELLULAR LOCALIZATION AND TRANSFORMATION OF EBF2 GENE IN TOMATO[J].Acta Agriculturae Nucleatae Sinica,2010,24(3):490-494. |
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| Authors: | WU Yu YANG Ying-wu DENG Wei LI Zheng-guo |
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| Institution: | 1.Genetic Engineering Research Center, Bio-Engineering College, Chongqing University,Chongqing 400030；
2.Key Lab of Functional Gene and New Regulation Technologies under Chongqing Municipal Education Commission, Chongqing 400030 |
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| Abstract: | In order to study the function of EBF2 cDNA in tomato, EBF2 full length gene from tomato fruit was amplified by RT-PCR. The transient expression of EBF2-GFP fusion protein in tobacco protoplast revealed its nuclear localization, and sense recombinant vector of pCambia1301-EBF2 which EBF2 gene driven by the constitutive promoter CaMV35S was constructed. Then the transgenic tomatoes cv.Micro Tom were obtained via Agrobacterium-mediated transformation and confirmed by PCR and GUS histochemical staining. |
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| Keywords: | EBF2 gene subcellular localization construction of expression vector transformation tomato |
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