Molecular cloning of invasion plasmid antigen (ipa) genes from Shigella flexneri: analysis of ipa gene products and genetic mapping.
作者： J M Buysse , C K Stover , E V Oaks , M Venkatesan , D J Kopecko
DOI: 10.1128/JB.169.6.2561-2569.1987
关键词:
摘要: Tn5-tagged invasion plasmid DNA (pWR110) from Shigella flexneri serotype 5 (strain M90T) was cloned into the expression vector lambda gt11. Recombinant phage (lambda gt11Sfl) expressing pWR110-encoded polypeptide antigens were identified by using rabbit antisera directed against S. M90T antigens. Antigens encoded gt11Sfl recombinant characterized reacting affinity-purified antibodies, eluted nitrocellulose-bound plaques of recombinants, with virulent, wild-type polypeptides in Western blot analyses. clones directing synthesis complete, truncated, and beta-galactosidase fusion versions three previously outer membrane (57-, 43-, 39-kilodalton [kDa] antigens) isolated. A fourth polypeptide, similar size to 57-kDa antigen (ca. 58 kDa) but unrelated as determined homology serological measurements, also identified. Southern analysis hybridized insert probes used construct a map genes (ipa) corresponding (ipaB), 43-kDa (ipaC), 39-kDa (ipaD) polypeptides. Genes ipaB, ipaC ipaD mapped contiguous 4.6-kilobase (kb) 1.0-kb HindIII fragments contained within larger (23-kb) BamHI fragment. The ipaH gene, which encodes 58-kDa did not or near ipaBCD gene cluster, suggesting distinct location on plasmid.
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