Helicobacter Pylori HpaA Recombinant Protein Expression And The Purification Of Inclusion Body

Objective: Use of genetic technology engineering and cloning hpaA gene expressed by Helicobacter pylori, the recombinant protein HpaA, through cutting the gel slices to purificate HpaA inclusion bodies, and identified its antigenicity and immunogenicity. Provide a simple economic approach of purification of H.pylori adhesion inclusion bodies.Methods: HpaA gene fragments was amplified by PCR from hpaA H.pylori DNA, and inserted the original expression vector pQE30, to transform into the receptors of DH5α. Cloning and sequence analysis in E. coli expression, SDS-PAGE analysis of protein molecular weight and form of expression, through the Western blot confirmed the antigenicity, and through the ELISA confirmed the immunogenicity. Take SDS-PAGE of the rHpaA into 4M sodium acetate show, cutting down the rHpaA can be used directly in mice, or be cutting down the purpose of dialysis equipment in the bag by the electric field, could be free rHpaA from a soluble protein, and for Western blot and ELISA test.Results: Recombinant expression plasmid pQE30-HpaA construction succeeded. The gene fragment inserted was 782 bp, and the gene bank of genetic homology HpaA of 97.0%. SDS-PAGE showed that expression of the relative molecular mass of 30kD, accounts for full expression of the total 32.3%, mainly exists in the inclusion body of the bacteria. ELISA and Western blot identification of the protein can be identified by anti-HpaA(+) serum.Conclusions: H.pylori hpaA gene expression cloning success, to obtain high-grade inclusion body as the main form of expression of the recombinant protein, confirmed the gel slice purification of rHpaA can still retain the original antigenicity and immunogenicity, and the traditional way of comparison, is simple and relatively economical and practical, rHpaA provide for the inclusion body of a new purification method.