The shufflon of IncI1 plasmids is rearranged constantly during different growth conditions

Publication date: Available online 15 March 2019Source: PlasmidAuthor(s): Michael S.M. Brouwer, Stephanie D. Jurburg, Frank Harders, Arie Kant, Dik J. Mevius, Adam P. Roberts, Alex BossersAbstractOne of the factors that can affect conjugation of IncI1 plasmids, amongst others, is the genetic region known as the shufflon. This multiple inversion system modifies the pilus tip proteins used during conjugation, thus affecting the affinity for different recipient cells. Although recombination is known to occur in in vitro conditions, little is known about the regulation and the extend of recombination that occurs. To measure the recombination of the shufflon, we have amplified the entire shufflon region and sequenced the amplicons using nanopore long-read sequencing. This method was effective to determine the order of the segments of the shufflon and allow for the analysis of the shufflon variants that are present in a heterogeneous pool of templates. Analysis was performed over different growth phases and after addition of cefotaxime. Furthermore, analysis was performed in different E. coli host cells to determine if recombination is likely to be influenced. Recombination of the shufflon was constantly ongoing in all conditions that were measured, although no differences in the amount of different shufflon variants or the rate at which novel variants were formed could be found. As previously reported, some variants were abundant in the population while others were scarce. This ...
Source: Plasmid - Category: Biotechnology Source Type: research