安徽农业科学Issue(13)：7645-7649,5.
重组蛋白PACAP-PTD的制备及其活性鉴定
Preparation and Activity Identification of Recombinant PACAP-PTD
摘要
Abstract
[Objective] This study aimed to prepare recombiriant protein PACAP-PTD and identify its activity. [Method] The gene that encodes fusion protein PACAP-PTD was cloned into the expression vector pKYB to construct rccombinant expression vector pKYB-PACAP-PTD, which was then transformed into E. coli ER2566. The fusion protein consisting of PACAP-FTD, intein and chitin was induced with IPTG to express. Finally, the target fusion protein PACAP-PTD was purified with purification system mediated by IMPACT (Intein Mediated Purification with an Affinity of chitin-binding Tag) , and its activity was identified. [Result] The molecular weight of target protein determined with laser time-of-flight mass spectrometry was consistent with the theoretical value. The recombinant protein PACAP-PTD could effectively cross the blood-brain barrier. [ Conclusion] The construction and preparation of the fusion protein PACAP-PTD not only lays foundation for further study on its biological function, but also improves the route of PA CAP administration, and thus expands its scope of application.关键词
PACAP-PTD/PACAP/内含肽/FITC/血脑屏障/原核表达Key words
PACAP-PTD/ PACAP/ Intein/ FTTC/ Blood-brain barrier/ Prokaryotic expression分类
农业科技引用本文复制引用
黄霖,余榕捷,李娟,刘晓飞,王静静..重组蛋白PACAP-PTD的制备及其活性鉴定[J].安徽农业科学,2012,(13):7645-7649,5.基金项目
东莞市科技计划项目(2008108101036). （2008108101036）
