Abstract
BackgroundSalmonella enterica serovar Enteritidis (S. Enteritidis) has emerged as one of the most important food-borne pathogens for humans. Lipopolysaccharide (LPS), as a component of the outer membrane, is responsible for the virulence and smooth-to-rough transition in S. Enteritidis. In this study, we screened S. Enteritidis signature-tagged transposon mutant library using monoclonal antibody against somatic O9 antigen (O9 MAb) and O9 factor rabbit antiserum to identify novel genes that are involved in smooth-to-rough transition.ResultsA total of 480 mutants were screened and one mutant with transposon insertion in rfbG gene had smooth-to-rough transition phenotype. In order to verify the role of rfbG gene, an rfbG insertion or deletion mutant was constructed using λ-Red recombination system. Phenotypic and biological analysis revealed that rfbG insertion or deletion mutants were similar to the wild-type strain in growth rate and biochemical properties, but the swimming motility was reduced. SE Slide Agglutination test and ELISA test showed that rfbG mutants do not stimulate animals to produce agglutinating antibody. In addition, the half-lethal dose (LD50) of the rfbG deletion mutant strain was 106.6 -fold higher than that of the parent strain in a mouse model when injected intraperitoneally.ConclusionsThese data indicate that the rfbG gene is involved in smooth-to-rough transition, swimming motility and virulence of S. Enteritidis. Furthermore, somatic O-antigen antibody-based approach to screen signature-tagged transposon mutants is feasible to clarify LPS biosynthesis and to find suitable markers in DIVA-vaccine research.
Highlights
IntroductionEnteritidis) has emerged as one of the most important foodborne pathogens for humans
Enteritidis showing smooth-torough phenotype Out of 480 mutants screened, 1 potential mutant was not agglutinated with O9 Monoclonal antibody against somatic O9 antigen (MAb) or O9 factor rabbit antiserum
We identified some other genes that involved in smooth-to-rough transition, e.g., rfc, which has been used as a marker for distinguishing in Salmonella enterica vaccine research [22, 23]
Summary
Enteritidis) has emerged as one of the most important foodborne pathogens for humans. Lipopolysaccharide (LPS), as a component of the outer membrane, is responsible for the virulence and smooth-to-rough transition in S. Enteritidis signature-tagged transposon mutant library using monoclonal antibody against somatic O9 antigen (O9 MAb) and O9 factor rabbit antiserum to identify novel genes that are involved in smooth-to-rough transition. Enteritidis, SE) has emerged as one of the most important foodborne pathogens for humans, with poultry meat and eggs being the most common sources of human S. Enteritidis food-borne infections [1]. Young chicks showed high mortality rate when infected with S. Enteritidis usually leads to symptomless carriage, and is able to colonize the tissues of the ovary and oviduct of egg-
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