title: [Distribution of Salmonella paratyphi A pagC gene and immunoprotective effect of its recombinant expressed products].
abstract: To determine the distribution and sequence conservation of pagC gene in Salmonella paratyphi A isolates, and the immunogenicity and immunoprotection of its recombinant expression products (rPagC).
  
  
  
  The distribution of pagC gene in Salmonella paratyphi A isolates and its sequence conservation were examined by PCR and sequencing. A prokaryotic expression system of pagC gene was constructed and the expressed rPagC was extracted by Ni-NTA affinity chromatography. SDS-PAGE and Bio-Rad Gel Image Analyzer were applied to examine the expression and yield of rPagC. The antigenicity and immunoreactivity of rPagC were detected by immunodiffusion test, ELISA and Western Blot assay. The immunoprotective effect of rPagC against infection of Salmonella paratyphi A in mice was determined, while the agglutinative effect of sera from rPagC-immunized mice was measured by micro-Widal's test.
  
  
  
  All the Salmonella paratyphi A isolates tested had the pagC gene, the similarity of nucleotide and amino acid sequences was 99.1 %-100 % and 98.4 %-100 %, respectively. The constructed prokaryotic expression system expressed rPagC with high efficiency. The rPagC immunized rabbit produced a high level antibody and it also combined with antiserum against whole cell of S. paratyphi A to generate a positive Western hybridization signal. ELISA results indicated that 97.1 % (66/68) paratyphoid patients infected with Salmonella paratyphi A were positive for rPagC antibody in their serum specimens. When mice were immunized with 100 μg or 200 μg rPagC, the immunoprotective rates were 73.3 % (11/15) or 86.7 % (13/15), respectively. The sera from rPagC-immunized mice offered 1:10-1:40 agglutination titers with the H antigens of Salmonella paratyphi A and Salmonella typhi.
  
  
  
  PagC gene has an extensive distribution in Salmonella paratyphi A isolates. rPagC can be used as the candidate antigen in genetic engineering vaccine due to its fine immunogenicity and powerful immunoprotective effect.
authors.affiliation: School of Laboratory Medical Science and Life Science, Wenzhou Medical College, Wenzhou 325035, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Zhang
authors.forename: Xinli
authors.initials: X
authors.lastname: Fan
authors.forename: Yumei
authors.initials: Y
authors.lastname: Ge
authors.forename: Jie
authors.initials: J
authors.lastname: Yan
authors.forename: Aihua
authors.initials: A
authors.lastname: Sun
mesh_terms: D000372:Agglutination Tests; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D001428:Bacterial Vaccines; D008565:Membrane Proteins / Q000235:genetics* / Q000276:immunology; D051379:Mice; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012482:Salmonella paratyphi A / Q000235:genetics* / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D008565:Membrane Proteins; D011994:Recombinant Proteins; C066766:pagC protein, Salmonella typhimurium