Analysis of hemolytic determinants of plasmid pHly185 by Tn5 mutagenesis

A nonconjugative hemolysin plasmid, pHly185, was isolated from Escherichia coli 185. Tn5 mutagenesis produced nonhemolytic derivatives that either expressed no hemolytic activity or did not secrete activity. These Tn5 insertions were located in three contiguous EcoRI fragments. Insertions inactivating hemolysin structural gene(s) were identified on all three EcoRI fragments. Mutations affecting the secretory function were clustered in one portion of a single EcoRI fragment.