Recombinant expression and immunological characterization of hemolysin coregulated protein 1, Burkholderia pseudomallei type Ⅵ secretion system protein

Abstract

Objective To express recombinant hemolysin coregulated protein 1 (Hcp1) of type Ⅵ secretion system (T6SS) of Burkholderia pseudomallei and identify its immunological properties. Methods The recombinant Hcp1 protein was expressed by using DNA recombination technology through pET-28a expression system in E.coli BL21(DE3), and then purified by affinity chromatography. Then 6-week-old BALB/c mice were immunized with the obtained Hcp1 via intraperitoneal injection, the serum samples were collected to detect the titers of anti-Hcp1 specific antibodies by ELISA. Meanwhile, the immune response against antigen-specific antibodies was evaluated by Western blotting and immunofluorescence assay. Results The recombinant Hcp1 protein was successfully obtained, with a purity of 95%. The obtained recombinant protein stimulated the antisera of good specificity and high titer (1: 512 000), with good immunogenicity and immunoreactivity. Conclusion The recombinant protein Hcp1 and its antisera are successfully prepared.

Keywords

• burkholderia pseudomallei
• type ⅵ secretion system protein
• hemolysin coregulated protein 1
• recombinant expression