title: [Expression of Proteus mirabilis polyphosphate kinase and preparation of its polyclonal antibodies].
abstract: To express and purify polyphosphate kinase (PPK) from Proteus mirabilis and prepare the polyclonal antibody against PPK.
  
  
  
  The antigenicity and hydrophobicity of PPK were analyzed using software. The N-terminal conservative sequence containing 309 amino acids was selected as the target peptide, and its corresponding gene sequence with modification based on prokaryotic cells-preferred codon was synthesized and inserted into plasmid pET28b(+). The constructed recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and induced with IPTG. The expressed fusion protein was purified using Ni-affinity chromatography. The purified protein was injected along with adjuvant in rabbits to prepare the polyclonal antibodies against PPK.
  
  
  
  PPK fusion protein expressed by E. coli was purified successfully using Ni-affinity chromatography. ELISA result demonstrated that the harvested rabbit anti-sera against PPK had a high titer of 1:512 000, and Western blotting showed a good specificity of the antibody, which can be used further study of the role of PPK in the pathogenesis of Proteus mirabilis infection.
authors.affiliation: Department of Clinical Laboratory, Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, China.E-mail: pl_206@126.com.
authors.forename: Liang
authors.initials: L
authors.lastname: Peng
authors.forename: Jing-Yi
authors.initials: JY
authors.lastname: Ou
authors.forename: Jia-Yun
authors.initials: JY
authors.lastname: Pan
authors.forename: Cong
authors.initials: C
authors.lastname: Deng
authors.forename: Jing-Hong
authors.initials: JH
authors.lastname: Chen
authors.forename: Hong
authors.initials: H
authors.lastname: Cao
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D004926:Escherichia coli; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000235:genetics / Q000276:immunology*; D011513:Proteus mirabilis / Q000201:enzymology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000906:Antibodies; D011993:Recombinant Fusion Proteins; D017856:Phosphotransferases (Phosphate Group Acceptor); C022424:polyphosphate kinase