[Preparation of polyclonal antibody against recombinant sigma⁵⁴ protein from Helicobacter pylori]

Objective To express and purify recombinant sigma⁵⁴ protein of Helicobacter pylori, and prepare polyclonal antibody against sigma⁵⁴ protein. Methods Intact rpoN gene fragment was amplified from Helicobacter pylori (H. pylori) 26695 genome, and then cloned into prokaryotic expression vector pTriEx^TM-4 with ⁶His 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 sigma⁵⁴ 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 sigma⁵⁴ protein. Conclusion The study successfully expressed and purified sigma⁵⁴ protein, and obtained its polyclonal antibody.