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BMC Microbiol. 2017 Mar 3;17(1):48. doi: 10.1186/s12866-017-0951-4.

Signature-tagged mutagenesis screening revealed a novel smooth-to-rough transition determinant of Salmonella enterica serovar Enteritidis.

Jiao Y1,2, Guo R1,2, Tang P1,2, Kang X1,2, Yin J1,2, Wu K1,2, Geng S1,2, Li Q1,2, Sun J2,3, Xu X2, Zhou X4, Gan J1,2, Jiao X1,2, Liu X5,6, Pan Z7,8.

Author information

1
Jiangsu Key Laboratory of Zoonosis, Yangzhou University, 48 East Wenhui Road, Yangzhou, Jiangsu, 225009, China.
2
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, 225009, China.
3
Division of Gastroenterology and Hepatology Department of Medicine, University of Illinois at Chicago, 840 S Wood Street, Room 704 CSB, 60612, Chicago, IL, USA.
4
Pathobiology and Veterinary Science, College of Agriculture, Health and Natural Resources, University of Connecticut, 61 North Eagleville Road, Unit-3089, Mansfield, CT, USA.
5
Jiangsu Key Laboratory of Zoonosis, Yangzhou University, 48 East Wenhui Road, Yangzhou, Jiangsu, 225009, China. xfliu@yzu.edu.cn.
6
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, 225009, China. xfliu@yzu.edu.cn.
7
Jiangsu Key Laboratory of Zoonosis, Yangzhou University, 48 East Wenhui Road, Yangzhou, Jiangsu, 225009, China. zmpan@yzu.edu.cn.
8
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, 225009, China. zmpan@yzu.edu.cn.

Abstract

BACKGROUND:

Salmonella 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.

RESULTS:

A 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.

CONCLUSIONS:

These 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.

KEYWORDS:

O9 MAb; S. Enteritidis; Signature-tagged mutagenesis (STM); Smooth-to-rough transition; rfbG gene

PMID:
28253852
PMCID:
PMC5335844
DOI:
10.1186/s12866-017-0951-4
[Indexed for MEDLINE]
Free PMC Article

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