Supplementary Material for: Revealing the Latent Mobilization Capability of the Staphylococcal Bacteriocinogenic Plasmid pRJ9

Plasmid pRJ9 is a non-self-mobilizable bacteriocinogenic plasmid from <i>Staphylococcus aureus</i>. Despite this feature, DNA sequencing and RT-PCR experiments showed that it presents a Mob region with three genes <i>(mobCAB)</i>, transcribed as an operon. In silico analysis of the Mob proteins encoded by pRJ9 showed that they present all the conserved functional features reported until present as being essential for plasmid mobilization. Moreover, they showed a high identity to Mob proteins encoded by mobilizable plasmids from <i>Staphylococcus</i> spp., especially to those encoded by plasmid pRJ6, which presents four <i>mob</i> genes <i>(mobCDAB)</i>. A putative <i>oriT </i>region was also found upstream of the pRJ9 <i>mob</i> operon. pRJ9 could only be successfully mobilized by pGO1 when pRJ6 was present in the same strain. Further experiments showed that the pRJ9 <i>oriT</i> can be recognized by the pRJ6 Mob proteins, confirming its functionality. As pRJ9 does not possess a <i>mobD </i>gene while pRJ6 does, the absence of this gene was believed to be responsible for its lack of mobilization. However, conjugation experiments with a donor strain carrying also <i>mobD</i> cloned into an <i>S. aureus </i>vector showed that pRJ9 does not become mobilized even in the presence of the protein MobD encoded by pRJ6. Therefore, the reasons for pRJ9 failure to be mobilized are presently unknown.