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To escape from the host immune system, the meningitis\u2010causing bacteria must survive and multiply in the bloodstream, relying on specific virulence factors such as capsules. Therefore, it is essential to study the genes involved in capsule biosynthesis in\n                    <jats:italic>R. anatipestifer<\/jats:italic>\n                    . In this study, we successfully constructed gene deletion mutants \u03943820 and \u03943830, targeting the\n                    <jats:italic>GE296_RS03820<\/jats:italic>\n                    and\n                    <jats:italic>GE296_RS03830<\/jats:italic>\n                    genes, respectively, using the RA\u2010LZ01 strain as the parental strain. The growth kinetics analysis revealed that these two genes contribute to bacterial growth. Transmission and scanning electron microscopy (TEM and SEM) and silver staining showed that \u03943820 and \u03943830 produced the altered capsules and compounds of capsular polysaccharides (CPSs). Serum resistance test showed the mutants also exhibited reduced C3b deposition and decreased resistance serum killing. In\u00a0vivo, \u03943820 and \u03943830 exhibited markedly declining capacity to cross the blood\u2013brain barrier, compared to RA\u2010LZ01. These findings indicate that the\n                    <jats:italic>GE296_RS03820<\/jats:italic>\n                    and\n                    <jats:italic>GE296_RS03830<\/jats:italic>\n                    genes are involved in CPSs biosynthesis and play a key role in the pathogenicity of\n                    <jats:italic>R. anatipestifer.<\/jats:italic>\n                    Furthermore, \u03943820 and \u03943830 mutants presented a tendency toward higher survival rates from RA\u2010LZ01 challenge in\u00a0vivo. Additionally, sera from ducklings immunized with the mutants showed cross\u2010immunoreactivity with different serotypes of\n                    <jats:italic>R. anatipestifer<\/jats:italic>\n                    , including 1, 2, 7 and 10. Western blot and SDS\u2010PAGE assays revealed that the altered CPSs of \u03943820 and \u03943830 resulted in the exposure of some conserved proteins playing the key role in the cross\u2010immunoreactivity. Our study clearly demonstrated that the\n                    <jats:italic>GE296_RS03820<\/jats:italic>\n                    and\n                    <jats:italic>GE296_RS03830<\/jats:italic>\n                    genes are involved in CPS biosynthesis in\n                    <jats:italic>R. anatipestifer<\/jats:italic>\n                    and the capsule is a target for attenuation in vaccine development.\n                  <\/jats:p>","DOI":"10.1096\/fj.202302694rr","type":"journal-article","created":{"date-parts":[[2024,7,2]],"date-time":"2024-07-02T12:01:09Z","timestamp":1719921669000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["The\n                    <i>GE296_RS03820<\/i>\n                    and\n                    <i>GE296_RS03830<\/i>\n                    genes are involved in capsular polysaccharide biosynthesis in\n                    <i>Riemerella anatipestifer<\/i>"],"prefix":"10.1096","volume":"38","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-7228-5605","authenticated-orcid":false,"given":"Xiaoni","family":"Wu","sequence":"first","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences  Lanzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-0976-2607","authenticated-orcid":false,"given":"Jiazhen","family":"Ge","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences  Lanzhou 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