Study on the cloning and expression and the immunogenicity of Helicobacter pylori adhesin AlpA gene / 中华消化杂志

ABSTRACT

Objective To construct a recombinant strain which expresses adhesin AlpA of Helicobacter pylori(H.pylori) and to study the immunogenicity of adhesin AlpA. Methods The adhesin AlpA DNA was amplified by PCR and inserted into the prokaryotie expression vector pET-22b(+)and expressed in the BL21(DE3) E.coli strain. Adhesin AlpA immunogenicity was studied by Western blot test. Results DNA sequence analysis showed that the sequence of adhesin AlpA DNA was almost the same as that of the published by GenBank. The adhesin AlpA recombinant protein accounted for 31.9% of the total bacterial protein. Western blot analysis of rAlpA confirmed that it could be specially recognized by serum from rabbit immunized with AlpA itself and H.pylori infected patients. Conclusion Cloning and high-expression of adhesin AlpA and primary confirmation of its immunogenicity lay the foundation for H.pylori gene engineering vaccine preparation and adhering mechanism research.