Signature-tagged mutagenesis screening revealed the role of lipopolysaccharide biosynthesis gene rfbH in smooth-to-rough transition in Salmonella Enteritidis

Yang Jiao; Zemiao Xia; Xiaohui Zhou; Yaxin Guo; Rongxian Guo; Xilong Kang; Kaiyue Wu; Jun Sun; Xiulong Xu; Xinan Jiao; Zhiming Pan; Xiufan Liu

doi:10.1016/j.micres.2018.05.001

Signature-tagged mutagenesis screening revealed the role of lipopolysaccharide biosynthesis gene rfbH in smooth-to-rough transition in Salmonella Enteritidis

Microbiol Res. 2018 Jul-Aug:212-213:75-79. doi: 10.1016/j.micres.2018.05.001. Epub 2018 May 5.

Authors

Yang Jiao¹, Zemiao Xia¹, Xiaohui Zhou², Yaxin Guo¹, Rongxian Guo¹, Xilong Kang¹, Kaiyue Wu¹, Jun Sun³, Xiulong Xu¹, Xinan Jiao¹, Zhiming Pan⁴, Xiufan Liu⁵

Affiliations

¹ Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, MOA, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, Jiangsu, 225009, China.
² Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, MOA, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Pathobiology and Veterinary Science, College of Agriculture, Health and Natural Resources, University of Connecticut, 61 North Eagleville Road, Unit-3089, Mansfield, CT, USA.
³ Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Division of Gastroenterology and Hepatology, College of Medicine, University of Illinois at Chicago, Chicago, IL, 60612, USA.
⁴ Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, MOA, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, Jiangsu, 225009, China. Electronic address: zmpan@yzu.edu.cn.
⁵ Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, MOA, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, Jiangsu, 225009, China. Electronic address: xfliu@yzu.edu.cn.

PMID: 29853170
DOI: 10.1016/j.micres.2018.05.001

Abstract

Salmonella enterica serovar Enteritidis (S. Enteritidis, SE) is a major cause of foodborne diseases for humans. The completeness of the O-chain antigen of Lipopolysaccharide (LPS) determines whether a S. Enteritidis strain is smooth or rough. However, genes that are involved in the synthesis of LPS and rough-smooth variation are not completely understood. In this study, we used monoclonal antibody against O-antigens (O₉ mAb) to identify novel factors that are involved in LPS synthesis and rough variation in S. Enteritidis by using signature-tagged mutagenesis (STM) technique. Our results showed that transposon insertion in the gene rfbH led to different LPS phenotype, auto-aggregation characteristic, motility and resistance to environmental stress compared with SE wild-type strain C50041. In addition, sera tests showed that rfbH mutant does not elicit specific antibodies against O-antigens in vaccinated animals. Taken together, the S. Enteritidis rfbH gene is implicated in LPS biosynthesis, rough variation, sera distinguishable reaction, motility and stress resistance. The rfbH mutant strain could be potentially used as a distinguishable vaccine or a live vector to deliver drugs and antibodies in vivo.

Keywords: Rough strain; Salmonella Enteritidis; Signature-tagged mutagenesis (STM); rfbH gene.

MeSH terms

Agglutination Tests
Animals
Antibodies, Bacterial
Antibodies, Monoclonal
Antigens, Bacterial
Bacterial Proteins / genetics*
Humans
Lipopolysaccharides / biosynthesis*
Lipopolysaccharides / genetics*
Mutagenesis*
Mutagenesis, Insertional
Mutagenicity Tests
O Antigens / genetics
O Antigens / immunology
Salmonella Infections, Animal / prevention & control
Salmonella enteritidis / genetics*
Salmonella enteritidis / pathogenicity
Virulence / genetics

Substances

Antibodies, Bacterial
Antibodies, Monoclonal
Antigens, Bacterial
Bacterial Proteins
Lipopolysaccharides
O Antigens
rfb protein, Bacteria
vaccin