title: [Preparation of polyclonal antibody against recombinant sigma54protein from Helicobacter pylori].
abstract: Objective To express and purify recombinant sigma54protein of Helicobacter pylori, and prepare polyclonal antibody against sigma54protein. Methods Intact rpoN gene fragment was amplified from Helicobacter pylori (H. pylori) 26695 genome, and then cloned into prokaryotic expression vector pTriExTM-4 with6His 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 sigma54protein 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 sigma54protein. Conclusion The study successfully expressed and purified sigma54protein, and obtained its polyclonal antibody.
authors.affiliation: Department of Clinical Laboratory, First Affiliated Hospital, Dalian Medical University, Dalian 116011, China. *Corresponding author, E-mail: liushuang498@163.com.
authors.forename: Shuang
authors.initials: S
authors.lastname: Liu
mesh_terms: D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D016480:Helicobacter pylori / Q000276:immunology*; D051877:RNA Polymerase Sigma 54 / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology*
chemical_list: D000906:Antibodies; D011993:Recombinant Fusion Proteins; D051877:RNA Polymerase Sigma 54