In addition, the 35-kb HPI of Yersinia enterocolitica could be mobilised [51] when a modified RP4 plasmid was used as a shuttle vector during the transfer experiments. Several cases

of plasmid mobilisation as a major mechanism check details for horizontal gene transfer of PAIs have been described [42–44]. With the PAI II536 construct used in this study, we were able to transfer this ~107-kb DNA region in the presence of the unmodified RP4 plasmid and thereby demonstrated that PAI II536 is mobilisable, but not self-transmissible. To increase the stability of the large PAI II536-specific CI and thus the transfer frequency, we also integrated an origin of replication into this PAI. In this respect, our model construct is artificial, but exhibits similar features of some ICEs including the HPIECOR31.

In the latter case, the origin of replication seems to be inactivated by insertion of an IS630 homologue [33]. This may explain why HPIECOR31 is not transferable although CI formation of this island was shown in the same study. Whereas plasmids replicate autonomously, ICEs are generally thought to be incapable of autonomous replication. Instead, their replication depends on that of host chromosome [52]. Some ICE and ICE-like elements, however, have been reported to be capable of autonomous replication [53–57]. In the light F plasmid-mediated mobilization of the HPI [13], it would, nevertheless, also be interesting to analyse in the future if a PAI II536 construct, which is not a self-replicating entity, but

only carries an oriT, could be mobilized upon provision of the appropriate conjugative buy CYC202 machinery in trans on a plasmid. The primary aim of our study was to demonstrate the transferability of a large archetypal island of UPEC strain 536 as this PAI can be excised selleck chemical site-specifically from the chromosome by its cognate integrase. On the other hand, we also tested conditions which may affect the transfer of an excised circular PAI intermediate. The frequency of PAI transfer in the mobilisation experiments was low (between 10-8 and 10-9). We postulate that the efficiency of PAI II536 transfer depends on several factors including the growth temperature, integrase activity, the size, and the chromosomal or episomal state of the PAI. In spite of the large size of PAI II536, complete transfer occurred at a high rate. 93.1% of the transconjugants received the complete 107-kb PAI II536 construct. The activity of the PAI-encoded integrase can contribute to the transfer efficiency by affecting the PAI FG-4592 molecular weight excision as well as the integration frequency. The remobilisation efficiency was three log scales higher with a stable episomal CI compared to an integrated PAI, indicating that a more active integrase may increase the chance of transfer by frequent induction of PAI-excision from the chromosome (Table 1). PAI II536 transfer rates at 20°C and 37°C were not significantly different. Besides the gut, E.