Prokaryotic Expression and Identification of PADRE-TGFβ1 Fusion Protein

To construct the prokaryotic expression PADRE-TGFβ1 plasmid and analyze the immunogenicity of fusion protein,the codon-optimized gene sequence of PADRE-Linker-TGFβ1 was synthesized and inserted into pET28 plasmid,then transformed into E.coli BL21 competent cells.The recombinant protein PADRE-TGFβ1 was induced by 1 mmol/L IPTG and the protein was detected by SDS-PAGE and Western blot.The expressed protein had a MW of 19.87 ku which consistent with the expected protein MW.Western blot analysis showed that the protein could specifically combine with TGFβ1 antibody.The results showed that pET28-PADRE-TGFβ1 prokaryotic expression vector was constructed successfully and the induced PADRE-TGFβ1 fusion protein has the immunogenicity of TGFβ1.This would lay a foundation for preparation and application of TGFβ1 autologous vaccine.