[Objective] This study aimed to prepare recombinant protein PACAP-PTD and measure its activity. [Method] The gene that encodes fusion protein PACAP-PTD was cloned into the expression vector pKYB to construct recombina...[Objective] This study aimed to prepare recombinant protein PACAP-PTD and measure its activity. [Method] The gene that encodes fusion protein PACAP-PTD was cloned into the expression vector pKYB to construct recombinant expression vector pKYB-PACAP-PTD, which was then transformed into E. coli ER2566. The fusion protein consisting of PACAP-PTD, intein and chitin was expressed under the induction of IPTG. Finally, the target fusion protein PACAP-PTD was purified with IMPACT system （ Intein Mediated Purification with an Affinity of chitin-binding Tag）, and its activities to cross blood-brain barrier and to promote cell proliferation were measured. [ Result~ The molecular weight of the fusion protein PACAP-PTD determined with laser time-of-flight mass spectrometry was con- sistent with the theoretical value. In addition, the protein could effectively cross the blood-brain barrier and promote cell proliferation as well. [ 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 PACAP administration, and thus expands its scope of application.展开更多
The effects of oligodeoxynucleotide （ODN） on the conformation of basic fibroblast growth factor （bFGF） were studied by spectral method. The results showed that ODN destabilized the protein.
基金Supported by Science and Technology Program of Dongguan City ( 2008108101036)
文摘[Objective] This study aimed to prepare recombinant protein PACAP-PTD and measure its activity. [Method] The gene that encodes fusion protein PACAP-PTD was cloned into the expression vector pKYB to construct recombinant expression vector pKYB-PACAP-PTD, which was then transformed into E. coli ER2566. The fusion protein consisting of PACAP-PTD, intein and chitin was expressed under the induction of IPTG. Finally, the target fusion protein PACAP-PTD was purified with IMPACT system （ Intein Mediated Purification with an Affinity of chitin-binding Tag）, and its activities to cross blood-brain barrier and to promote cell proliferation were measured. [ Result~ The molecular weight of the fusion protein PACAP-PTD determined with laser time-of-flight mass spectrometry was con- sistent with the theoretical value. In addition, the protein could effectively cross the blood-brain barrier and promote cell proliferation as well. [ 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 PACAP administration, and thus expands its scope of application.
文摘The effects of oligodeoxynucleotide （ODN） on the conformation of basic fibroblast growth factor （bFGF） were studied by spectral method. The results showed that ODN destabilized the protein.
