关键词: 人脂联素, 杆状病毒, 昆虫细胞, 表达

Abstract: Objective To investigate the expression of recombinant human adiponectin（ADPN） using baculovirus expression system in Sf9 insect cell line. Methods PCR method was used to amplify human ADPN gene,and then the gene was inserted into plasmid pFastBacl.Recombinant plasmid was transformed into E.coli DH10Bac which included a shuttle vector,Bacmid.Recombinant shuttle vector Bacmid-ADPN was obtained by site-specific transposition and transfect Sf9 cells. Identify expressed product by SDS-PAGE、Western blot. Results Sf9 cells infected by the recombinant baculovirus was observed distinct morphological changes. The Mw of recombinant protein was about 30 kD and Western blot proved that expressed protein could combine with ADPN polyclonal antibody.Conclusion Human ADPN gene was successfully expressed in Sf9 cells,and thus provided the basic material for studying its bioactivity and mechanism of action.

Key words: human adiponectin, baculovirus, insect cell, expression