[Preparation of polyclonal antibody against recombinant sigma 54 protein from Helicobacter pylori].

Objective To express and purify recombinant sigma protein of Helicobacter pylori, and prepare polyclonal antibody against sigma protein. Methods Intact rpoN gene fragment was amplified from Helicobacter pylori (H. pylori) 26695 genome, and then cloned into prokaryotic expression vector pTriEx-4 with His coding sequence. The recombinant plasmid was transformed into E.coli JM109DE. The recombinant protein with 6-His tag was expressed by IPTG induction and purified by His-Bind affinity columns. The purified protein supplemented with adjuvant was used to immunize rabbits to obtain the polyclonal antibody. Results Recombinant sigma protein was successfully expressed in E.coli JM109DE. SDS-PAGE that was used to detect the purified protein showed a single band which was consistent with the expected size of the recombinant protein. The polyclonal antibody exhibited a good recognition and specificity against sigma protein. Conclusion The study successfully expressed and purified sigma protein, and obtained its polyclonal antibody.