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. 2017 Feb 15;7:42724. doi: 10.1038/srep42724

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Copyright © 2017, The Author(s)

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(A) HpaM_Xoo (HpaM homologue in Xoo) and HpaM_Xoc (HpaM homologue in Xoc) are essential for type III secretion. Type III secretion signal sequence-gusA fusion reporter plasmid pGUSavrAC was introduced into Xoo and Xoc strains. The resulting recombinant strains were cultured in XOM2 medium for 12 h and the β-glucuronidase (GUS) activities in the culture (Total) and the cultural supernatant (Secreted) were determined. Values are the means ± standard deviation from three repeats. Left and right elements, GUS activities produced by pGUSavrAC in Xoo and Xoc strains, respectively. (B) The HpaM homologues HpaM_Xoo and HpaM_Xoc are also located in the outer membrane. The outer and inner membrane fraction proteins from Xoo and Xoc strains were prepared and 10 μg of each protein sample was separated by SDS-PAGE electrophoresis and transferred to a PVDF membrane. The presence of tested proteins was detected by anti-His₆ monoclonal antibody. The histidine sensor kinase HpaS and transcriptional regulator HpaR1 of Xcc were used as positive and negative controls. OM, outer membrane; IM, inner membrane.