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To provide a forward-genetics technology for finding virulent phenotype-related genes in these two pathogens, we constructed a novel temperature-sensitive suicide shuttle plasmid, pMar4s, which contains the\n                    <jats:italic>Himar1<\/jats:italic>\n                    system transposon, TnYLB-1, and the\n                    <jats:italic>Himar1<\/jats:italic>\n                    C9 transposase from pMarA and the\n                    <jats:italic>repTAs<\/jats:italic>\n                    temperature-sensitive fragment from pSET4s. The kanamycin (Kan) resistance gene was in the TnYLB-1 transposon. Temperature sensitivity and Kan resistance allowed the selection of mutant strains and construction of the mutant library. The SS2 and SEZ mutant libraries were successfully constructed using the pMar4s plasmid. Inverse-Polymerase Chain Reaction (Inverse-PCR) results revealed large variability in transposon insertion sites and that the library could be used for phenotype alteration screening. The thiamine biosynthesis gene\n                    <jats:italic>apbE<\/jats:italic>\n                    was screened for its influence on SS2 anti-phagocytosis; likewise, the\n                    <jats:italic>sagF<\/jats:italic>\n                    gene was identified to be a hemolytic activity-related gene in SEZ. pMar4s was suitable for mutant library construction, providing more information regarding SS2 and SEZ virulence factors and illustrating the pathogenesis of swine streptococcosis.\n                  <\/jats:p>","DOI":"10.1038\/srep27133","type":"journal-article","created":{"date-parts":[[2016,6,3]],"date-time":"2016-06-03T05:27:46Z","timestamp":1464931666000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["A novel suicide shuttle plasmid for Streptococcus suis serotype 2 and Streptococcus equi ssp. zooepidemicus gene mutation"],"prefix":"10.1038","volume":"6","author":[{"given":"Rui","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiqi","family":"Su","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huixing","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhe","family":"Ma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongjie","family":"Fan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,6,3]]},"reference":[{"key":"BFsrep27133_CR1","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1146\/annurev.genet.37.110801.142807","volume":"37","author":"F Hayes","year":"2003","unstructured":"Hayes, F. 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