The Combination of Bioinformatics With Biotechnology Would Expedite Vaccine Development Against Helicobactor. pylori

摘要

To screen the candidate gene for vaccine development, the published database of H.pylori strain 26695 and J99 was analyzed applying NCBI BLAST and software Omiga2.0 for genomic and proteomics mining. HP-napA was selected to be the target gene. HP-napA was cloned from a clinical H. Pylori strain MEL-HP27. After sequencing, the feature of HP-napA encode protein HP-NAP was predicted. Recombinant plasmid pMAL-c2x-napA was constructed and transformed into E.coli TB1. And the purified rHP-NAP was used as antigen to prepare rabbit anti-rHP-NAP IgG. The specific antibody against HP-NAP ir human sera was investigated with ELISA. rHP-NAP and antibody was used to test the inhibition of H.Pylori adherence to human gastric cancer cell line BGC823 in vitro. Our combined studies indicate that HP-napA was a highly conserved prokaryotic gene. HP-napA amplified from H. Pylori strain MEL- HP27, was a 435bp DNA fragment (Accession No AY366361). rHP-NAP could generate specific immunological reaction in rabbits. The ELISA assay showed that human anti-HP-NAP IgG existed in 85% of H.Pylori infected people and the positive rate of anti-NAP IgG in the adults involved in our investigation was 54%, similar to that of Urase 57%(P > 0.05). rHP-NAP and Anti-rHP-NAP IgG could partially inhibit H.Pylori adherence to gastric cancer cell line BGC823. This study demonstrate that biotechnology complements bioinformatics would expedite the process of vaccine development against H.Pylori.