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作 者 ：崔永兰;王鹏程;

期 刊 ：武汉植物学研究 2009年 27卷 2期 页码：216-220

关键词：拟南芥；叶绿体；转运肽；融合蛋白；

Keywords:Arabidopsis thaliana, Chloroplast, Transit peptide, Fusion protein,

摘 要 ：蛋白质的亚细胞定位对于深入了解该蛋白质所行使的生理功能具有重要意义。经生物信息学预测,两个拟南芥未知功能基因At4g16410与At1g18060编码蛋白含有叶绿体定位信息。我们分别克隆了这两个基因5’端长199bp与220bp的DNA片段,与绿色荧光蛋白(GFP)基因构建重组表达载体pMON530-cTP1-GFP与pMON530-cTP2-GFP,经农杆菌介导转化拟南芥。两种转基因植株经激光共聚焦显微镜观察,GFP荧光仅在叶绿体中观察到,表明所克隆的两段DNA序列编码的多肽能够将At4g16410与At1g18060编码蛋白质引导进入叶绿体,确定这两个蛋白质均为叶绿体蛋白质。

Abstract:The information of protein subcellular localization is important to our understanding of a protein function.The proteins encoded by both At4g16410 and At1g18060 were predicted to be the Arabidopsis chloroplast proteins.The 199 bp and 220 bp fragments of this two genes at the 5’ end were cloned and fused with GFP to construct the binary vectors pMON530-cTP1-GFP and pMON530-cTP2-GFP for genetic transformation,respectively.Observed by a confocal laser-scanning microscopy,green fluorescent signals were localized in chloroplasts in transgenic Arabidopsis plants suggesting the 199 bp and 220 bp fragments encoding peptides targeted to chloroplast.The results suggest that At4g16410 and At1g18060 encode two chloroplast proteins.

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