Qi Jing, Li Gui-Qin, Dong Zhen, Zhou Wei
College of Biology Science and Engineering, Hebei University of Economics and Business Shijiazhuang 050061, Hebei, China.
Hebei Women's Vocational College Shijiazhuang 050091, Hebei, China.
Am J Transl Res. 2016 Feb 15;8(2):698-704. eCollection 2016.
To explore the subcellular localization of Polyphenol oxidase (PPO) from Pyrus bretschneideri, the 1779 bp cDNA of PPO gene excluding the termination codon TAA was cloned and fused with GFP 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, suggesting that the Polyphenol oxidase from Pyrus bretschneideri was a chloroplast protein.
为探究砀山酥梨多酚氧化酶（PPO）的亚细胞定位，克隆了PPO基因不含终止密码子TAA的1779 bp cDNA，并将其与绿色荧光蛋白（GFP）融合构建双元载体pBI121 - PPO - GFP。然后，通过根癌农杆菌介导法将该双元载体转化到烟草中。利用共聚焦激光扫描显微镜观察发现，绿色荧光信号定位于转化后烟草细胞的叶绿体中，这表明砀山酥梨的多酚氧化酶是一种叶绿体蛋白。
