Localization of Plasmid Loci Necessary for the Entry of Shigella flexneri into HeLa Cells, and Characterization of One Locus Encoding Four Immunogenic Polypeptides

Abstract

We have previously cloned a 44kb fragment from the virulence plasmid of Shigella flexneri serotype 5 strain M90T which is capable of restoring invasiveness to an avirulent, plasmidless mutant. This report presents a genetic and physical analysis of Tn5 mutations in recombinant clone pHS4108. Tn5 mutagenesis allowed identification of at least five regions implicated in the entry phenotype. These regions were located on a 20 kb portion of pHS4108. Expression of the insertion mutants was studied by immunoblots using the serum of a convalescent monkey infected by S. flexneri 2a, which recognized four plasmid-associated polypeptides. We propose that the four immunogenic polypeptides, a, b, c and d, and encoded by an operon.