茶叶科学2012，Vol.32Issue(3)：269-275,7.
茶树PPO融合蛋白真核表达载体的构建与表达
Construction and Expression of Fusion Protein Eukaryotic Expression Vector of Polyphenol Oxidase Gene of Tea Plant (Camellia sinensis)
摘要
Abstract
The polyphenol oxidase (PPO) cds sequence was amplified by PCR from the genome of Yingshuang tea plant with the high fidelity polymerase pfu. The cds sequence was recombinanted into the eukaryotic expression vector pPICZa with the designed primer, and the fusion protein eukaryotic expression vector of tea polyphenol oxidase "pPICZa-PPO" was succeeded to be constructed, and then it was transformed into Pichia pasloris GS115. Therefore, several positive converters were screened. After those positive converters were induced by methanol, the target protein was successfully detected in the culture medium by using Western-Blotting method. The detection result of enzyme activity showed that the induced products had a relatively high enzyme activity and can correctly play their biological activity.关键词
茶树/多酚氧化酶/融合蛋白真核表达载体/巴斯德毕赤酵母/诱导表达Key words
Camellia sinensis, PPO, fusion protein eukaryotic expression vector, Pichia pastoris, inducible expression分类
农业科技引用本文复制引用
王乃栋,张丽霞,黄晓琴,韩晓阳,李智..茶树PPO融合蛋白真核表达载体的构建与表达[J].茶叶科学,2012,32(3):269-275,7.基金项目
高等学校博士学科点专项科研基金项目(20103702110002)、山东省自然科学基金面上项目(ZR2010CM026) （20103702110002）
