Expression and purification of human SNARE complex associated proteins
[Objective]The eukaryotic expression vectors of SNAP25,VAMP1 and Syntaxin 1a protein in human SNARE com-plex were constructed,and then expressed and purified in vitro.[Method]The recombinant plasmid was transformed into com-petent E.coli DH10Bac cells to construct the recombined bacmid and then transfected into Sf9 insect cells by liposome transfor-mation to produce recombinant virus,after that,the virus was amplified and infected Sf9 insect cells to express the target pro-tein.The obtained proteins were identified and purified by Western Blot.[Result]The sequencing results showed that the eu-karyotic expression vectors SNAP25,Syntaxin 1a and VAMP1 were successfully constructed.The specific bands of the three pro-teins were detected by Western Blot,and Syntaxin 1a was purified by His tag protein.[Conclusion]SNARE complex related proteins were successfully expressed by baculovirus expression system.
万方数据
维普
