| 摘要: |
| 将鸭梨PPO基因与绿色萤光蛋白GFP基因相融合共同进行遗传转化的方式,对鸭梨多酚氧化酶开展细胞定位研究。通过克隆该酶基因除终止密码子TAA外长度为1 779 bp的CDS序列,与绿色荧光蛋白基因重组构建了荧光表达载体pBI121-PPO-GFP,借助农杆菌转化烟草,转基因烟草叶片细胞经激光扫描共聚焦显微镜观察,绿色荧光蛋白荧光与叶绿体自发荧光相重合。结果表明鸭梨多酚氧化酶为叶绿体蛋白质。 |
| 关键词: 鸭梨 多酚氧化酶 绿色荧光蛋白 基因融合 遗传转化 亚细胞定位 |
| DOI：10.3969/j.issn.1000-3142.2014.03.016 |
| 分类号:Q942.6 |
| Fund project: |
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| Transformation of tobacco plants by Yali PPO-GFP fusion gene and observation of subcellular localization |
|
QI Jing1*, LI Gui-Qin1, DONG Zhen2, ZHOU Wei1
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1. College of Biology Science and Engineering, Hebei University of Economics and Business, Shijiazhuang
050061, China;2. Hebei Women's Vocational College, Shijiazhuang 050091, China
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| Abstract: |
| To explore the subcellular localization of Polyphenol oxidase(PPO)from Pyrus bretschneideri,the 1 779 bp cDNA of PPO gene was cloned and fused with GFP in frame to construct a binary vector pBI121-PPO-GFP. Then,the binary vector was transformed into Nicotiana tabacum by the tumefanciens-mediated method. Using confocal laser scanning microscopy,green fluorescent signals were localized in chloroplasts of the transformed Nicotiana tabacum cell. This results suggested that the Polyphenol oxidase from Pyrus bretschneideri was a chloroplast protein. |
| Key words: Yali polyphenol oxidase GFP gene fusion genetic transformation subcellular localization |
