Construction and Identification of the Eukaryotic Expression Vector of Chicken High Mobility Group B1 Protein

The purpose of this study was to make a eukaryotic expression construct of chicken high mobility group protein B1(chHMGB1). Then transient expression of the chHMGB1 gene in eukaryotic cells and identification of the protein expression level were carried out. The full length gene fragment of chicken high mobility group B1 protein was purified from PGEM-chHMGB1 plasmid by restriction enzyme digestion,which was ligated with pcDNA3.1(+)vector to construct pcDNA-chHMGB1 expression plasmid. Then the plasmid pcDNA-chHMGB1 was transfected into 293T cells for transient expression. chHMGB1 protein expression level was identified by Western blotting and indirect immunofluorescence assay. The results demonstrated that pcDNA-chHMGB1 recombinant construct was successfully made. The target protein,around 30 Kd,in the transient transfected 293T cells could be detected in the cytoplasma by western blot or IFA. Eukaryotic expression plasmid pcDNA-chHMGB1 were successfully constructed and were transiently transfected in eukaryotic cells,which provided useful materials for further biological function study.